In vitro transcription and screening kits for sgRNA
Our Guide-it in vitro transcription and screening kits for sgRNA include four kits for the simple production, cleanup, and evaluation of single guide RNAs (sgRNAs) for CRISPR/Cas9 studies. Each kit provides enough materials for 50 reactions. The Guide-it sgRNA In Vitro Transcription Kit (Cat. # 632635) is used to produce high yields of sgRNAs via in vitro transcription.
Our Guide-it in vitro transcription and screening kits for sgRNA include four kits for the simple production, cleanup, and evaluation of single guide RNAs (sgRNAs) for CRISPR/Cas9 studies. Each kit provides enough materials for 50 reactions. The Guide-it sgRNA In Vitro Transcription Kit (Cat. # 632635) is used to produce high yields of sgRNAs via in vitro transcription. This kit also includes the Guide-it IVT RNA Clean-Up Kit (Cat. # 632638), which is used to quickly purify sgRNA following in vitro transcription (IVT) reactions. Purified sgRNA from this kit is ready for direct use in transfection, electroporation, or in vitro cleavage assays. The Guide-it sgRNA Screening Kit (Cat. # 632639) enables you to test the efficacy of different single guide RNAs (sgRNAs) in vitro prior to using them in studies involving Cas9-mediated gene editing so that you don't waste time delivering ineffective sgRNAs to your cells. With this kit, a template containing a sgRNA target site is created by PCR, then combined with a sgRNA that you wish to test and recombinant Cas9 nuclease. The efficiency with which Cas9 nuclease cleaves the template can be measured using agarose gel electrophoresis. The Guide-it Complete sgRNA Screening System (Cat. # 632636) includes all of the kit components described above and is used to synthesize and test the efficacy of single guide RNAs (sgRNAs). The kit contains all the reagents needed to in vitro transcribe and purify sgRNAs, and test their efficacy against a PCR target fragment using recombinant Cas9 nuclease.
Overview
- Four kits for simple production, cleanup, and evaluation of the efficacy of sgRNA (single guide RNA) for Cas9-mediated gene editing studies
- In vitro transcribe and purify high yields of sgRNA with the Guide-it sgRNA In Vitro Transcription Kit (Cat. # 632635; includes the Guide-it IVT RNA Clean-Up Kit, Cat. # 632638) for transfection or in vitro screening
- Use the Guide-it IVT RNA Clean-Up Kit (Cat. # 632638) to quickly purify sgRNA after IVT reactions, for direct use in transfection, electroporation, or in vitro cleavage assays
- High-quality recombinant Cas9 protein included in the Guide-it sgRNA Screening Kit (Cat. # 632639)
- Best value: the Guide-it Complete sgRNA Screening System (Cat. # 632636) is a combination of both the Guide-it sgRNA In Vitro Transcription Kit (Cat. # 632635) and Guide-it sgRNA Screening Kit (Cat. # 632639), allowing you to both produce sgRNA and test its ability to cleave its putative target in vitro
More Information
Components
- Guide-it sgRNA In Vitro Transcription Kit (Cat. # 632635; 50 rxn)
- 1 each Guide-it sgRNA In Vitro Transcription Components v2 (Cat. # 632637; 50 rxn; Not sold separately)
- 50 µl Guide-it Scaffold Template (1 ng/µl)
- 350 µl Guide-it In Vitro Transcription Buffer
- 1 ml RNase-free Water
- 150 µl Guide-it T7 Polymerase Mix (33 U/µl)
- 625 µl PrimeSTAR Max Premix (2X)
- 100 µl Recombinant DNase I (RNase-free) (5 U/µl)
- 1 each Guide-it IVT RNA Clean-Up Kit (Cat. # 632638; 50 rxn)
- 1 each Guide-it sgRNA In Vitro Transcription Components v2 (Cat. # 632637; 50 rxn; Not sold separately)
- Guide-it sgRNA Clean-Up Kit (Cat. # 632638; 50 rxn)
- 13 ml RNase-free Water
- 3 ml IVT Binding Buffer
- 50 IVT RNA Clean-Up Spin Columns
- 2 x 6 ml IVT Wash Buffer
- 50 Collection Tubes (2 ml)
- Guide-it sgRNA Screening Kit (Cat. # 632639; 50 rxn)
- 25 µl Guide-it Recombinant Cas9 Nuclease (500 ng/µl)
- 50 µl 15X Cas9 Reaction Buffer
- 50 µl 15X BSA
- 5 µl Control sgRNA (50 ng/µl)
- 25 µl Control Fragment (20 ng/µl)
- 50 µl Terra PCR Direct Polymerase Mix (1.25 U/µl)
- 1.25 ml 2X Terra PCR Direct Buffer (with Mg2+, dNTP)
- 5 ml RNase-free Water
- 5 ml Extraction Buffer 1
- 0.5 ml Extraction Buffer 2
The Guide-it Complete sgRNA Screening System (Cat. # 632636; 50 rxn) is a combination of the Guide-it sgRNA In Vitro Transcription Kit (Cat. # 632635) and Guide-it sgRNA Screening Kit (Cat. # 632639) listed above.
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.
CRISPR/Cas9 information
Choosing sgRNA design tools
Browse a collection of sgRNA design tools for Cas9-based genome editing experiments.
Choosing a target sequence for CRISPR/Cas9 gene editing
Learn how to design sgRNA sequences for successful gene editing.
The CRISPR/Cas9 system for targeted genome editing
Overview of CRISPR/Cas9 system for genome editing.
CRISPR/Cas9 genome editing tools
An overview of tools available for each step in a successful genome editing workflow.
Gene editing technical notes
Delivery of Cas9 and sgRNA to mammalian cells using a variety of innovative tools.
SNP engineering application note
Learn about a simple assay for sensitive detection of single-nucleotide substitutions in bulk-edited or clonal cell populations.
CRISPR/Cas9 gesicles overview
Learn about Guide-it CRISPR/Cas9 Gesicle Production System components and workflow.
CRISPR library screening webinar
Watch this webinar to learn how you can perform genome-wide lentiviral sgRNA screens easily.
Choosing an HDR template format
Watch a webinar on how to choose the right HDR template for knockin experiments.
Guide-it SNP Screening Kit FAQs
Get answers to frequently asked questions and view a video explaining the enzymatic assay.
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