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  • Normalized Mate & Plate Libraries
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Yeast two-hybrid system

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Normalized yeast two-hybrid cDNA libraries—Mate & Plate

The Normalized Mate & Plate Libraries are high-complexity cDNA libraries cloned into a GAL4 AD vector and transformed into yeast strain Y187. These libraries significantly reduce the labor and time required to perform a yeast two-hybrid screen because library amplification and yeast transformation have already been done. Additionally, normalization selectively removes highly abundant transcripts from the libraries to enhance the representations of low-abundance and rare cDNAs.

The Normalized Mate & Plate Libraries are high-complexity cDNA libraries cloned into a GAL4 AD vector and transformed into yeast strain Y187. These libraries significantly reduce the labor and time required to perform a yeast two-hybrid screen because library amplification and yeast transformation have already been done. Additionally, normalization selectively removes highly abundant transcripts from the libraries to enhance the representations of low-abundance and rare cDNAs. This greatly reduces the possibility of obtaining false positives during screening.

Click to find additional yeast two-hybrid cDNA Mate and Plate libraries.

Is it really this easy?

Yes, there are just three simple steps:

  1. Make a bait strain expressing your protein of interest using the Matchmaker Gold Yeast Two-Hybrid System.
  2. Mix the bait strain overnight with a 1 ml vial of normalized library.
  3. Plate on selective medium.

What is a normalized library?

Normalization reduces the proportion of highly abundant transcripts in an cDNA pool. This means that many of the most highly abundant housekeeping genes are significantly reduced in copy number so you can screen fewer clones and be assured that your screens have included medium- and low-abundance clones. In short, you can screen a greater number of independent clones with less effort, and have a greater chance of detecting important interactions.

How are normalized libraries made?

We start by generating SMART-amplified cDNA, which we then normalize using duplex-specific nuclease (DSN) normalization (Shagin et al. 2002, Zhulidov et al. 2004). Briefly, the cDNA is denatured and reannealed, followed by brief digestion with an enzyme that specifically cleaves double-stranded DNA, because the more abundant cDNAs anneal more rapidly and thus are more likely to be destroyed by the enzyme. Finally, we check the efficiency of cDNA normalization by virtual Northern blot analysis and clone the normalized cDNA library into our prey vector, pGADT7-RecAB.

Universal libraries offer universal gene coverage

Our human and mouse universal libraries provide the broadest and most complete coverage of expressed genes. These normalized, all-purpose libraries were created from a diverse collection of whole tissues that were specifically chosen to represent the most expansive range of expressed genes (Zhulidov et al. 2004). Combining “across-the-board” gene representation with the enrichment of low-copy-number cDNAs, our Mate & Plate Universal (Normalized) Libraries offer the greatest capacity for identifying novel and genuine binding partners for your protein of interest.

Why use normalized universal libraries?

Many of the most meaningful interactions in nature occur between appropriately localized, but weakly expressed proteins. These types of interactions may not be readily detectable in a traditional, single-tissue library. Universal libraries that are also normalized offer an effective solution to this problem because they represent a balanced array of transcripts covering the broadest possible range of expressed genes. Thus, your screens are not limited to identifying interactions between proteins that are highly expressed in a single tissue.

 More  Less
Cat. # Product Size License Quantity Details
630487 Mate & Plate™ Library - Universal Arabidopsis (Normalized) 5 x 1 mL USD $1352.00

This yeast two-hybrid library was constructed from mRNA isolated from 11 Arabidopsis tissues, mixed in equal quantities and transformed into yeast strain Y187. The cDNA was normalized prior to library construction to reduce the copy number of abundant cDNAs derived from highly represented mRNAs, thereby increasing the representation of low copy number transcripts. The normalization process combines a Duplex-Specific Nuclease (DSN) treatment and SMART technology, reduces the number of clones that must be screened in your yeast two-hybrid assay, and facilitates the identification and characterization of novel protein-protein interactions. The library was transformed into yeast strain Y187 and can be readily mated to a MATa GAL4 reporter strain, such as AH109 or Y2HGold, for screening.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Reduction in abundance of highly expressed gene transcripts following cDNA normalization

Reduction in abundance of highly expressed gene transcripts following cDNA normalization
Reduction in abundance of highly expressed gene transcripts following cDNA normalization. cDNA normalization reduces the abundance of highly expressed gene transcripts. Data are shown for genes from mixed tissues, before and after normalization, which exhibit greater than 10,000 Mean Fluorescence Units (MFU), representing over 7,000 genes. Approximately 3,300 genes show a significant reduction in intensity, and thus abundance, following normalization. Due to the large volume of data obtained, the median MFU was plotted for groups of 100 genes before and after normalization.

Back

630487: Mate & Plate Library - Universal Arabidopsis (Normalized)

630487: Mate & Plate Library - Universal Arabidopsis (Normalized)

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.

Back

Matchmaker Gold reporter genes

Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630486 Mate & Plate™ Library - Human Brain (Normalized) 5 x 1 mL USD $1352.00

This yeast two-hybrid library was constructed from mRNA isolated from human brain tissue and transformed into yeast strain Y187. The cDNA was normalized prior to library construction to reduce the copy number of abundant cDNAs derived from highly represented mRNAs, thereby increasing the representation of low copy number transcripts. The normalization process combines a Duplex-Specific Nuclease (DSN) treatment and SMART technology, reduces the number of clones that must be screened in your yeast two-hybrid assay, and facilitates the identification and characterization of novel protein-protein interactions. The library was transformed into yeast strain Y187 and can be readily mated to a MATa GAL4 reporter strain, such as AH109 or Y2HGold, for screening.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

Back

Use of pretransformed libraries simplifies screening

Use of pretransformed libraries simplifies screening
Use of pretransformed libraries simplifies screening.

Back

DSN-Normalization reduces the amount of highly abundant transcripts

DSN-Normalization reduces the amount of highly abundant transcripts
DSN-Normalization reduces the amount of highly abundant transcripts. Normalized (Lanes N) and non-normalized (Lanes C) Human Universal cDNA samples (PCR products) were electrophoresed on a 1.5% agarose gel and transferred to Hybond-N membrane. PCR-amplified probes of GAPDH and beta-actin were labeled with 32P-dATP and hybridized to the membrane.

Back

630486: Mate & Plate Library - Human Brain (Normalized)

630486: Mate & Plate Library - Human Brain (Normalized)

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.

Back

Matchmaker Gold reporter genes

Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630485 Mate & Plate™ Library - Universal Drosophila (Normalized) 5 x 1 mL USD $1352.00

This yeast two-hybrid library was constructed from mRNA isolated from Drosophila melanogaster and transformed into yeast strain Y187. The cDNA was normalized prior to library construction to reduce the copy number of abundant cDNAs derived from highly represented mRNAs, thereby increasing the representation of low copy number transcripts. The normalization process combines a Duplex-Specific Nuclease (DSN) treatment and SMART technology, reduces the number of clones that must be screened in your yeast two-hybrid assay, and facilitates the identification and characterization of novel protein-protein interactions.The library was transformed into yeast strain Y187 and can be readily mated to a MATa GAL4 reporter strain, such as AH109 or Y2HGold, for screening.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

Back

Use of pretransformed libraries simplifies screening

Use of pretransformed libraries simplifies screening
Use of pretransformed libraries simplifies screening.

Back

Reduction in abundance of highly expressed gene transcripts following cDNA normalization

Reduction in abundance of highly expressed gene transcripts following cDNA normalization
Reduction in abundance of highly expressed gene transcripts following cDNA normalization. cDNA normalization reduces the abundance of highly expressed gene transcripts. Data are shown for genes from mixed tissues, before and after normalization, which exhibit greater than 10,000 Mean Fluorescence Units (MFU), representing over 7,000 genes. Approximately 3,300 genes show a significant reduction in intensity, and thus abundance, following normalization. Due to the large volume of data obtained, the median MFU was plotted for groups of 100 genes before and after normalization.

Back

DSN-Normalization reduces the amount of highly abundant transcripts

DSN-Normalization reduces the amount of highly abundant transcripts
DSN-Normalization reduces the amount of highly abundant transcripts. Normalized (Lanes N) and non-normalized (Lanes C) Human Universal cDNA samples (PCR products) were electrophoresed on a 1.5% agarose gel and transferred to Hybond-N membrane. PCR-amplified probes of GAPDH and beta-actin were labeled with 32P-dATP and hybridized to the membrane.

Back

630485: Mate & Plate Library - Universal Drosophila (Normalized)

630485: Mate & Plate Library - Universal Drosophila (Normalized)

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.

Back

Matchmaker Gold reporter genes

Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630484 Mate & Plate™ Library - Mouse Embryonic Stem Cell (Normalized) 5 x 1 mL USD $1352.00

This yeast two-hybrid library was constructed from mRNA isolated from mouse embryonic stem cells (E14TG2A cell line) and transformed into yeast strain Y187. The cDNA was normalized prior to library construction to reduce the copy number of abundant cDNAs derived from highly represented mRNAs, thereby increasing the representation of low-copy-number transcripts. The normalization process combines a Duplex-Specific Nuclease (DSN) treatment and SMART technology, reduces the number of clones that must be screened in your yeast two-hybrid assay, and facilitates the identification and characterization of novel protein-protein interactions.

The library was transformed into yeast strain Y187 and can be readily mated to a MATa GAL4 reporter strain, such as AH109 or Y2HGold, for screening.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

Back

Use of pretransformed libraries simplifies screening

Use of pretransformed libraries simplifies screening
Use of pretransformed libraries simplifies screening.

Back

Reduction in abundance of highly expressed gene transcripts following cDNA normalization

Reduction in abundance of highly expressed gene transcripts following cDNA normalization
Reduction in abundance of highly expressed gene transcripts following cDNA normalization. cDNA normalization reduces the abundance of highly expressed gene transcripts. Data are shown for genes from mixed tissues, before and after normalization, which exhibit greater than 10,000 Mean Fluorescence Units (MFU), representing over 7,000 genes. Approximately 3,300 genes show a significant reduction in intensity, and thus abundance, following normalization. Due to the large volume of data obtained, the median MFU was plotted for groups of 100 genes before and after normalization.

Back

DSN-Normalization reduces the amount of highly abundant transcripts

DSN-Normalization reduces the amount of highly abundant transcripts
DSN-Normalization reduces the amount of highly abundant transcripts. Normalized (Lanes N) and non-normalized (Lanes C) Human Universal cDNA samples (PCR products) were electrophoresed on a 1.5% agarose gel and transferred to Hybond-N membrane. PCR-amplified probes of GAPDH and beta-actin were labeled with 32P-dATP and hybridized to the membrane.

Back

630484: Mate & Plate Library - Mouse Embryonic Stem Cell (Normalized)

630484: Mate & Plate Library - Mouse Embryonic Stem Cell (Normalized)

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.

Back

Matchmaker Gold reporter genes

Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630483 Mate & Plate™ Library - Universal Mouse (Normalized) 5 x 1 mL USD $1352.00

This yeast two-hybrid library was constructed from mouse cDNA that had been previously normalized to preferentially remove abundant cDNAs derived from high-copy-number mRNAs. The normalization process incorporates a Duplex-Specific Nuclease (DSN) treatment and SMART technology, and increases the representation of low-copy-number transcripts in the library. This reduces the number of clones that must be screened to identify positive interactions, and facilitates the identification and characterization of novel protein-protein interactions.

A universal mouse cDNA library transformed into yeast strain Y187. The library can be readily mated to a MATa GAL4 reporter strain, such as AH109 or Y2HGold.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

Back

Use of pretransformed libraries simplifies screening

Use of pretransformed libraries simplifies screening
Use of pretransformed libraries simplifies screening.

Back

Reduction in abundance of highly expressed gene transcripts following cDNA normalization

Reduction in abundance of highly expressed gene transcripts following cDNA normalization
Reduction in abundance of highly expressed gene transcripts following cDNA normalization. cDNA normalization reduces the abundance of highly expressed gene transcripts. Data are shown for genes from mixed tissues, before and after normalization, which exhibit greater than 10,000 Mean Fluorescence Units (MFU), representing over 7,000 genes. Approximately 3,300 genes show a significant reduction in intensity, and thus abundance, following normalization. Due to the large volume of data obtained, the median MFU was plotted for groups of 100 genes before and after normalization.

Back

DSN-Normalization reduces the amount of highly abundant transcripts

DSN-Normalization reduces the amount of highly abundant transcripts
DSN-Normalization reduces the amount of highly abundant transcripts. Normalized (Lanes N) and non-normalized (Lanes C) Human Universal cDNA samples (PCR products) were electrophoresed on a 1.5% agarose gel and transferred to Hybond-N membrane. PCR-amplified probes of GAPDH and beta-actin were labeled with 32P-dATP and hybridized to the membrane.

Back

630483: Mate & Plate Library - Universal Mouse (Normalized)

630483: Mate & Plate Library - Universal Mouse (Normalized)

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.

Back

Matchmaker Gold reporter genes

Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630482 Mate & Plate™ Library - Universal Mouse (Normalized) 2 x 1 mL USD $665.00

This yeast two-hybrid library was constructed from mouse cDNA that had been previously normalized to preferentially remove abundant cDNAs derived from high-copy-number mRNAs. The normalization process incorporates a Duplex-Specific Nuclease (DSN) treatment and SMART technology, and increases the representation of low-copy-number transcripts in the library. This reduces the number of clones that must be screened to identify positive interactions, and facilitates the identification and characterization of novel protein-protein interactions.

A universal mouse cDNA library transformed into yeast strain Y187. The library can be readily mated to a MATa GAL4 reporter strain, such as AH109 or Y2HGold.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

Back

Use of pretransformed libraries simplifies screening

Use of pretransformed libraries simplifies screening
Use of pretransformed libraries simplifies screening.

Back

Reduction in abundance of highly expressed gene transcripts following cDNA normalization

Reduction in abundance of highly expressed gene transcripts following cDNA normalization
Reduction in abundance of highly expressed gene transcripts following cDNA normalization. cDNA normalization reduces the abundance of highly expressed gene transcripts. Data are shown for genes from mixed tissues, before and after normalization, which exhibit greater than 10,000 Mean Fluorescence Units (MFU), representing over 7,000 genes. Approximately 3,300 genes show a significant reduction in intensity, and thus abundance, following normalization. Due to the large volume of data obtained, the median MFU was plotted for groups of 100 genes before and after normalization.

Back

DSN-Normalization reduces the amount of highly abundant transcripts

DSN-Normalization reduces the amount of highly abundant transcripts
DSN-Normalization reduces the amount of highly abundant transcripts. Normalized (Lanes N) and non-normalized (Lanes C) Human Universal cDNA samples (PCR products) were electrophoresed on a 1.5% agarose gel and transferred to Hybond-N membrane. PCR-amplified probes of GAPDH and beta-actin were labeled with 32P-dATP and hybridized to the membrane.

Back

630482: Mate & Plate Library - Universal Mouse (Normalized)

630482: Mate & Plate Library - Universal Mouse (Normalized)

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.

Back

Matchmaker Gold reporter genes

Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630481 Mate & Plate™ Library - Universal Human (Normalized) 2 x 1 mL USD $665.00

This yeast two-hybrid library was constructed from human cDNA that had been previously normalized to preferentially remove abundant cDNAs derived from high-copy-number mRNAs. The normalization process incorporates a Duplex-Specific Nuclease (DSN) treatment and SMART technology, and increases the representation of low-copy-number transcripts in the library. This reduces the number of clones that must be screened to identify positive interactions, and facilitates the identification and characterization of novel protein-protein interactions.

A universal human cDNA library transformed into yeast strain Y187. The library can be readily mated to a MATa GAL4 reporter strain, such as AH109 or Y2HGold.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

Back

Use of pretransformed libraries simplifies screening

Use of pretransformed libraries simplifies screening
Use of pretransformed libraries simplifies screening.

Back

Reduction in abundance of highly expressed gene transcripts following cDNA normalization

Reduction in abundance of highly expressed gene transcripts following cDNA normalization
Reduction in abundance of highly expressed gene transcripts following cDNA normalization. cDNA normalization reduces the abundance of highly expressed gene transcripts. Data are shown for genes from mixed tissues, before and after normalization, which exhibit greater than 10,000 Mean Fluorescence Units (MFU), representing over 7,000 genes. Approximately 3,300 genes show a significant reduction in intensity, and thus abundance, following normalization. Due to the large volume of data obtained, the median MFU was plotted for groups of 100 genes before and after normalization.

Back

DSN-Normalization reduces the amount of highly abundant transcripts

DSN-Normalization reduces the amount of highly abundant transcripts
DSN-Normalization reduces the amount of highly abundant transcripts. Normalized (Lanes N) and non-normalized (Lanes C) Human Universal cDNA samples (PCR products) were electrophoresed on a 1.5% agarose gel and transferred to Hybond-N membrane. PCR-amplified probes of GAPDH and beta-actin were labeled with 32P-dATP and hybridized to the membrane.

Back

630481: Mate & Plate Library - Universal Human (Normalized)

630481: Mate & Plate Library - Universal Human (Normalized)

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.

Back

Matchmaker Gold reporter genes

Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630480 Mate & Plate™ Library - Universal Human (Normalized) 5 x 1 mL USD $1352.00

This yeast two-hybrid library was constructed from human cDNA that had been previously normalized to preferentially remove abundant cDNAs derived from high-copy-number mRNAs. The normalization process incorporates a Duplex-Specific Nuclease (DSN) treatment and SMART technology, and increases the representation of low-copy-number transcripts in the library. This reduces the number of clones that must be screened to identify positive interactions, and facilitates the identification and characterization of novel protein-protein interactions.

A universal human cDNA library transformed into yeast strain Y187. The library can be readily mated to a MATa GAL4 reporter strain, such as AH109 or Y2HGold.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

Back

Use of pretransformed libraries simplifies screening

Use of pretransformed libraries simplifies screening
Use of pretransformed libraries simplifies screening.

Back

Reduction in abundance of highly expressed gene transcripts following cDNA normalization

Reduction in abundance of highly expressed gene transcripts following cDNA normalization
Reduction in abundance of highly expressed gene transcripts following cDNA normalization. cDNA normalization reduces the abundance of highly expressed gene transcripts. Data are shown for genes from mixed tissues, before and after normalization, which exhibit greater than 10,000 Mean Fluorescence Units (MFU), representing over 7,000 genes. Approximately 3,300 genes show a significant reduction in intensity, and thus abundance, following normalization. Due to the large volume of data obtained, the median MFU was plotted for groups of 100 genes before and after normalization.

Back

DSN-Normalization reduces the amount of highly abundant transcripts

DSN-Normalization reduces the amount of highly abundant transcripts
DSN-Normalization reduces the amount of highly abundant transcripts. Normalized (Lanes N) and non-normalized (Lanes C) Human Universal cDNA samples (PCR products) were electrophoresed on a 1.5% agarose gel and transferred to Hybond-N membrane. PCR-amplified probes of GAPDH and beta-actin were labeled with 32P-dATP and hybridized to the membrane.

Back

630480: Mate & Plate Library - Universal Human (Normalized)

630480: Mate & Plate Library - Universal Human (Normalized)

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.

Back

Matchmaker Gold reporter genes

Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System
630479 Mate & Plate™ Library - HeLa S3 (Normalized) 5 x 1 mL USD $1352.00

This yeast two-hybrid library was constructed from HeLa S3 cDNA that had been previously normalized to preferentially remove abundant cDNAs derived from high-copy-number mRNAs. The normalization process incorporates a Duplex-Specific Nuclease (DSN) treatment and SMART technology, and increases the representation of low-copy-number transcripts in the library. This reduces the number of clones that must be screened to identify positive interactions, and facilitates the identification and characterization of novel protein-protein interactions.

HeLa S3 cDNA library transformed into yeast strain Y187. The library can be readily mated to a MATaGAL4 reporter strain, such as AH109 or Y2HGold.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

Back

Use of pretransformed libraries simplifies screening

Use of pretransformed libraries simplifies screening
Use of pretransformed libraries simplifies screening.

Back

Reduction in abundance of highly expressed gene transcripts following cDNA normalization

Reduction in abundance of highly expressed gene transcripts following cDNA normalization
Reduction in abundance of highly expressed gene transcripts following cDNA normalization. cDNA normalization reduces the abundance of highly expressed gene transcripts. Data are shown for genes from mixed tissues, before and after normalization, which exhibit greater than 10,000 Mean Fluorescence Units (MFU), representing over 7,000 genes. Approximately 3,300 genes show a significant reduction in intensity, and thus abundance, following normalization. Due to the large volume of data obtained, the median MFU was plotted for groups of 100 genes before and after normalization.

Back

DSN-Normalization reduces the amount of highly abundant transcripts

DSN-Normalization reduces the amount of highly abundant transcripts
DSN-Normalization reduces the amount of highly abundant transcripts. Normalized (Lanes N) and non-normalized (Lanes C) Human Universal cDNA samples (PCR products) were electrophoresed on a 1.5% agarose gel and transferred to Hybond-N membrane. PCR-amplified probes of GAPDH and beta-actin were labeled with 32P-dATP and hybridized to the membrane.

Back

630479: Mate & Plate Library - HeLa S3 (Normalized)

630479: Mate & Plate Library - HeLa S3 (Normalized)

Required Products

Cat. # Product Size Price License Quantity Details
630489 Matchmaker® Gold Yeast Two-Hybrid System Each USD $1059.00

Complete GAL4-based two-hybrid system to detect protein interactions. Yeast strain Y2HGold facilitates the elimination of many false positives that arise when screening an activation domain (AD) fusion library, because it contains four separate reporter genes that can be used to select for protein interactions. The system is also characterized by low background due to the Aureobasidin A resistance marker.

Documents Components You May Also Like Image Data

Back

The yeast two-hybrid principle

The yeast two-hybrid principle
The yeast two-hybrid principle. A bait protein interacts with the GAL4 recognition sequence (or promoter) upstream of a reporter gene. Transcription of the reporter is activated when a prey protein containing the GAL4 transcriptional activation domain interacts with the bait.

Back

Matchmaker Gold reporter genes

Matchmaker Gold reporter genes
Matchmaker Gold reporter genes. Yeast strain Y2HGold expresses 4 genes from 3 separate GAL4-responsive promoters in response to protein-protein interactions.

Back

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency

Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency
Four reporters give the Matchmaker Gold Yeast Two-Hybrid System its high stringency. Interacting bait and prey fusion proteins drive the expression of four different reporters from three different GAL4-responsive promoters (M1, G1, and G2), which are stably integrated in the genome of the reporter strain, Y2H Gold. Aureobasidin A (AbA) resistance and the two auxotrophic reporters for histidine and adenine biosynthesis confer growth selection in the presence of AbA and on histidine- and adenine-deficient media, while the a-galactosidase reporter produces blue colonies in the presence of X-alpha-Gal.

Back

630489: Matchmaker Gold Yeast Two-Hybrid System

630489: Matchmaker Gold Yeast Two-Hybrid System

Overview

  • By far the easiest way to screen a library for protein-protein interactions
  • Screen fewer colonies, detect more interactions
  • Universal libraries—broadest gene representation
  • No more searching for needles in a haystack

More Information

Applications

  • Yeast two-hybrid library screening

References

Shagin, D. A. et al. A novel method for SNP detection using a new duplex-specific nuclease from crab hepatopancreas. Genome Res. 12, 1935–1942 (2002).         

Zhulidov, P. A. et al. Simple cDNA normalization using kamchatka crab duplex-specific nuclease. Nucleic Acids Res. 32, e37 (2004).

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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