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  • ‹ Back to CRISPR-Cas9
  • Long ssDNA for knockins
  • Knockin screening kit
  • Genome-wide sgRNA library system
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  • Mutation detection kits
  • In vitro transcription and screening kits
  • Cas9-sgRNA gesicle production
  • Cas9 antibodies
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Guide-it Recombinant Cas9 protein Recombinant Cas9 protein
Long ssDNA for CRISPR/Cas9 gene knockin experiments Long ssDNA for CRISPR/Cas9 gene knockin
Home › Products › Gene function › Gene editing › CRISPR-Cas9 › In vitro transcription and screening kits

Gene editing

  • CRISPR-Cas9
    • Long ssDNA for knockins
    • Knockin screening kit
    • Genome-wide sgRNA library system
    • Recombinant Cas9 protein
    • GMP recombinant Cas9
    • Mutation detection kits
    • In vitro transcription and screening kits
    • Cas9-sgRNA gesicle production
    • Cas9 antibodies
    • Plasmid systems
    • Lentiviral systems
    • AAV systems
    • RNA transfection
    • Genotype confirmation kit
    • Indel identification kit
  • Cre recombinase
    • AAV2-Cre recombinase
    • Cre Recombinase Gesicles
    • Cre recombinase RLPs
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Guide-it Recombinant Cas9 protein Recombinant Cas9 protein
Long ssDNA for CRISPR/Cas9 gene knockin experiments Long ssDNA for CRISPR/Cas9 gene knockin

In vitro transcription and screening kits for sgRNA

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.

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Cat. # Product Size Price License Quantity Details
632638 Guide-it™ IVT RNA Clean-Up Kit 50 Rxns USD $426.00

The Guide-it IVT RNA Clean-Up Kit is included as part of the Guide-it In Vitro Transcription Kit and is used to purify sgRNA transcribed using the Guide-it In Vitro Transcription Kit.

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 Image Data

Back

Production of sgRNA by in vitro transcription and testing for cleavage using recombinant Cas9 protein

Production of sgRNA by in vitro transcription and testing for cleavage using recombinant Cas9 protein
Schematic highlighting the key steps for using Guide-it kits for the production of high amounts of single guide RNA (sgRNA) and screening for target site cleavage efficacy using recombinant Cas9 protein.

Back

sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System

sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System
sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System. Panel A. 20 µl of each sgRNA in vitro transcription (IVT) reaction was incubated for 4 hr at 37°C. The yield for each transcribed sgRNA was quantified using a NanoDrop spectrophotometer. Panel B. Each sgRNA IVT reaction was a run on an Agilent Bioanalyzer to assay for sample quality.

Back

sgRNA quality produced via in vitro transcription

sgRNA quality produced via in vitro transcription
sgRNA quality produced via in vitro transcription. sgRNAs produced using the Guide-it sgRNA In Vitro Transcription Kit were compared with sgRNAs produced using a competitor’s kit. The Guide-it kit produced a high-quality, single band for each reaction, while the competitor’s kit showed unwanted byproducts.

Back

Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs

Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs
Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs.
632639 Guide-it™ sgRNA Screening Kit 50 Rxns USD $475.00

License Statement

ID Number  
391 LIMITED USE LABEL LICENSE: RESEARCH USE ONLY Notice to Purchaser: This product is the subject to a license granted to Takara Bio USA, Inc. and its Affiliates from Caribou Biosciences, Inc., and this product is transferred to the end-user purchaser (“Purchaser”) subject to a “Limited Use Label License” conveying to the Purchaser a limited, non-transferable right to use the product, solely as provided to Purchaser, together with (i) progeny or derivatives of the product generated by the Purchaser (including but not limited to cells), and (ii) biological material extracted or derived from the product or its corresponding progeny or derivatives (including but not limited to cells) (collectively, the product, and (i) and (ii) are referred to as (“Material”) only to perform internal research for the sole benefit of the Purchaser. The Purchaser cannot sell or otherwise transfer Material to a third party or otherwise use the Material for any Excluded Use. “Excluded Use” means any and all: (a) commercial activity including, but not limited to, any use in manufacturing (including but not limited to cell line development for purposes of bioproduction), product testing, or quality control; (b) preclinical or clinical testing or other activity directed toward the submission of data to the U.S. Food and Drug Administration, or any other regulatory agency in any country or jurisdiction where the active agent in such studies comprises the Material; (c) use to provide a service, information, or data to a third party; (d) use for human or animal therapeutic, diagnostic, or prophylactic purposes or as a product for therapeutics, diagnostics, or prophylaxis; (e) activity in an agricultural field trial or any activity directed toward the submission of data to the U.S. Department of Agriculture or any other agriculture regulatory agency; (f) high throughput screening drug discovery purposes (i.e., the screening of more than 10,000 experiments per day) as well as scale-up production activities for commercialization; (g) modification of human germline, including editing of human embryo genomes (with the sole exception of editing human embryonic stem (ES) cell lines for research purposes) or reproductive cells; (h) self-editing; and/or (i) stimulation of biased inheritance of a particular gene or trait or set of genes or traits (“gene drive”). It is the Purchaser’s responsibility to use the Material in accordance with all applicable laws and regulations. For information on obtaining additional rights, including commercial rights, please contact licensing@cariboubio.com or Caribou Biosciences, Inc., 2929 7th Street, Suite 105, Berkeley, CA 94710 USA, Attn: Licensing.

The Guide-it sgRNA Screening Kit contains PCR reagents to amplify genomic DNA from your target cells and recombinant Cas9 for the in vitro screening of your sgRNA to help you pick the best guide sequences before moving to experiments in living cells or animals. It is combined with the Guide-it In VItro Transcription kit as part of the Guide-it Complete sgRNA Screening System.

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 Image Data

Back

Production of sgRNA by in vitro transcription and testing for cleavage using recombinant Cas9 protein

Production of sgRNA by in vitro transcription and testing for cleavage using recombinant Cas9 protein
Schematic highlighting the key steps for using Guide-it kits for the production of high amounts of single guide RNA (sgRNA) and screening for target site cleavage efficacy using recombinant Cas9 protein.

Back

sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System

sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System
sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System. Panel A. 20 µl of each sgRNA in vitro transcription (IVT) reaction was incubated for 4 hr at 37°C. The yield for each transcribed sgRNA was quantified using a NanoDrop spectrophotometer. Panel B. Each sgRNA IVT reaction was a run on an Agilent Bioanalyzer to assay for sample quality.

Back

sgRNA quality produced via in vitro transcription

sgRNA quality produced via in vitro transcription
sgRNA quality produced via in vitro transcription. sgRNAs produced using the Guide-it sgRNA In Vitro Transcription Kit were compared with sgRNAs produced using a competitor’s kit. The Guide-it kit produced a high-quality, single band for each reaction, while the competitor’s kit showed unwanted byproducts.

Back

Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs

Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs
Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs.

Back

632639: Guide-it sgRNA Screening Kit

632639: Guide-it sgRNA Screening Kit
632636 Guide-it™ Complete sgRNA Screening System 50 Rxns USD $1056.00

License Statement

ID Number  
M54 This product is covered by the claims of U.S. Patent Nos. 7,704,713 and its foreign counterparts. 
272 This product (“Product”) and its use, is the subject of U.S. Patents 8,697,359 and 8,771,945 and pending U.S. Patent applications. The purchase of the Product conveys to the buyer the non-transferable right to use Product(s) purchased from Takara Bio USA, Inc. or its Affiliates, and any progeny, modification or derivative of a Product, or any cell or animal made or modified through use of a Product, or any progeny, modification or derivative of such cell or animal (“Related Material”), solely for research conducted by the buyer in accordance with all of the following requirements. No right is given to use this Product or Related Material for any other purpose, including, but not limited to, use in drugs, in vitro diagnostic purposes, therapeutics, or in humans. (i) The buyer shall not sell or otherwise transfer Products (including without limitation any material that contains a Licensed Product in whole or part) or any Related Material to any other person or entity, or use Products or any Related Material to perform services for the benefit of any other person or entity, (ii) the buyer shall use only the purchased amount of the Products and components of the Products, and shall use any Related Material, only for its internal research and not for (a) the practice, performance or provision of any method, process or service, or (b) the manufacture, sale, use, distribution, disposition or importing of any product, in each case (a) or (b) for consideration, or on any other commercial basis (“Commercial Purpose”), (iii) the buyer shall use Licensed Products and any Related Material in compliance with all applicable laws and regulations, including without limitation applicable human health and animal welfare laws and regulations, and (iv) the buyer shall indemnify, defend and hold harmless MIT, Harvard and The Broad and their current and former trustees, directors, officers, faculty, affiliated investigators, students, employees, and agents and their respective successors, heirs and assigns (“Indemnitees”), against any liability, damage, loss, or expense (including without limitation reasonable attorneys’ fees and expenses) incurred by or imposed upon any of the Indemnitees in connection with any claims, suits, investigations, actions, demands or judgments arising out of or related to the exercise of any rights granted to the buyer, or any breach of the rights granted hereunder by the buyer.
391 LIMITED USE LABEL LICENSE: RESEARCH USE ONLY Notice to Purchaser: This product is the subject to a license granted to Takara Bio USA, Inc. and its Affiliates from Caribou Biosciences, Inc., and this product is transferred to the end-user purchaser (“Purchaser”) subject to a “Limited Use Label License” conveying to the Purchaser a limited, non-transferable right to use the product, solely as provided to Purchaser, together with (i) progeny or derivatives of the product generated by the Purchaser (including but not limited to cells), and (ii) biological material extracted or derived from the product or its corresponding progeny or derivatives (including but not limited to cells) (collectively, the product, and (i) and (ii) are referred to as (“Material”) only to perform internal research for the sole benefit of the Purchaser. The Purchaser cannot sell or otherwise transfer Material to a third party or otherwise use the Material for any Excluded Use. “Excluded Use” means any and all: (a) commercial activity including, but not limited to, any use in manufacturing (including but not limited to cell line development for purposes of bioproduction), product testing, or quality control; (b) preclinical or clinical testing or other activity directed toward the submission of data to the U.S. Food and Drug Administration, or any other regulatory agency in any country or jurisdiction where the active agent in such studies comprises the Material; (c) use to provide a service, information, or data to a third party; (d) use for human or animal therapeutic, diagnostic, or prophylactic purposes or as a product for therapeutics, diagnostics, or prophylaxis; (e) activity in an agricultural field trial or any activity directed toward the submission of data to the U.S. Department of Agriculture or any other agriculture regulatory agency; (f) high throughput screening drug discovery purposes (i.e., the screening of more than 10,000 experiments per day) as well as scale-up production activities for commercialization; (g) modification of human germline, including editing of human embryo genomes (with the sole exception of editing human embryonic stem (ES) cell lines for research purposes) or reproductive cells; (h) self-editing; and/or (i) stimulation of biased inheritance of a particular gene or trait or set of genes or traits (“gene drive”). It is the Purchaser’s responsibility to use the Material in accordance with all applicable laws and regulations. For information on obtaining additional rights, including commercial rights, please contact licensing@cariboubio.com or Caribou Biosciences, Inc., 2929 7th Street, Suite 105, Berkeley, CA 94710 USA, Attn: Licensing.
396 Sigma-Aldrich CRISPR Use License Agreement This Product and its use are the subject of one or more of the following issued patents and patent applications: Australia Patent Nos. 2013355214; 2017204031; and 2018229489; Canada Patent Nos. 2,891,347 and 2,977,152; China Patent No. CN105142669; European Patent Nos. EP 2 928 496 B1; EP 3 138 910 B1, 3 138 911 B1, EP 3 138 912 B1, EP 3 360 964 B1, EP 3 363 902 B1; Israel Patent No. IL238856; Singapore Patent No. 11201503824S; South Korea Patent Nos. 10-1844123 and 10-2006880; and U.S. Patent Application Serial Nos. 15/188,911; 15/188,924; 15/188,927; 15/188,931; and 15/456,204 (the “Patent Rights”). The purchase of this Product conveys to you (the “Buyer”) the NON-TRANSFERABLE right to use the Product for Licensed Research Use (see definition below) subject to the conditions set out in this License Agreement. 1. “Licensed Research Use” means any use for research purposes, except: (i) Buyer may not sell or otherwise transfer the Product (including without limitation any material that contains the Product in whole or part) or any Related Material to any other third party (except that you may transfer the Product, or any Related Material to a bona fide collaborator or contract research organization), or use the Products or any Related Material to perform services for the benefit of any other third party; (ii) Buyer may use only the purchased amount of the Product and components of the Product, and shall use any Related Material, only for your internal research within the Field, and not for any Commercial Purposes; (iii) Buyer shall use the Product and any Related Material in compliance with all applicable laws and regulations, including without limitation applicable human health and animal welfare laws and regulations; and (iv) the Buyer shall indemnify, defend, and hold harmless SIGMA and their current and former directors, officers, employees and agents, and their respective successors, heirs and assigns (the “Indemnities”) against any liability, damage, loss, or expense (including without limitation reasonable attorneys’ fees and expenses) incurred by or imposed upon any of the Indemnitees in connection with any claims, suits, investigations, actions, demands or judgments arising out of or related to the exercise of any rights granted to the Buyer hereunder or any breach of this License Agreement by such Buyer. 2. For purposes of Section 1 above, the following definitions shall apply: “Commercial Purposes” means (a) the practice, performance or provision of any method, process or service, or (b) the manufacture, sale, use, distribution, disposition or importing of any product, in each case (a) or (b) for consideration, or on any other commercial basis. “Field” means use as a research tool for research purposes; provided, however, that notwithstanding the foregoing, the Field shall expressly exclude (a) any in vivo and ex vivo human or clinical use, including, without limitation, any administration into humans or any diagnostic or prognostic use, (b) the creation of transgenic rodent models and/or derivatives thereof (including, but not limited to, rodents’ cells and rodents’ organs) by for-profit entities, (c) any in vivo veterinary or livestock use, or non-research agricultural use, or (d) use as a testing service, therapeutic or diagnostic for humans or animals. “Related Materials” means any progeny, modification or derivative of a Product. 3. Your right to use the Product will terminate immediately if you fail to comply with these terms and conditions. You shall, upon such termination of your rights, destroy all Product, Related Materials, and components thereof in your control, and notify SIGMA of such in writing. For information on purchasing a license to this Product for purposes other than Licensed Research Use, contact your local SIGMA sales representative, or call +1 800-325-3010.

The Guide-it Complete sgRNA Screening System includes everything needed for the simple production, cleanup, and evaluation of single guide RNAs (sgRNAs) for CRISPR/Cas9 studies, including PCR reagents for amplifying your genomic target and recombinant Cas9 for in vitro analysis of the transcribed sgRNA.

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 Image Data

Back

Production of sgRNA by in vitro transcription and testing for cleavage using recombinant Cas9 protein

Production of sgRNA by in vitro transcription and testing for cleavage using recombinant Cas9 protein
Schematic highlighting the key steps for using Guide-it kits for the production of high amounts of single guide RNA (sgRNA) and screening for target site cleavage efficacy using recombinant Cas9 protein.

Back

sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System

sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System
sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System. Panel A. 20 µl of each sgRNA in vitro transcription (IVT) reaction was incubated for 4 hr at 37°C. The yield for each transcribed sgRNA was quantified using a NanoDrop spectrophotometer. Panel B. Each sgRNA IVT reaction was a run on an Agilent Bioanalyzer to assay for sample quality.

Back

sgRNA quality produced via in vitro transcription

sgRNA quality produced via in vitro transcription
sgRNA quality produced via in vitro transcription. sgRNAs produced using the Guide-it sgRNA In Vitro Transcription Kit were compared with sgRNAs produced using a competitor’s kit. The Guide-it kit produced a high-quality, single band for each reaction, while the competitor’s kit showed unwanted byproducts.

Back

Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs

Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs
Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs.
632635 Guide-it™ sgRNA In Vitro Transcription Kit 50 Rxns USD $881.00

License Statement

ID Number  
M54 This product is covered by the claims of U.S. Patent Nos. 7,704,713 and its foreign counterparts. 
272 This product (“Product”) and its use, is the subject of U.S. Patents 8,697,359 and 8,771,945 and pending U.S. Patent applications. The purchase of the Product conveys to the buyer the non-transferable right to use Product(s) purchased from Takara Bio USA, Inc. or its Affiliates, and any progeny, modification or derivative of a Product, or any cell or animal made or modified through use of a Product, or any progeny, modification or derivative of such cell or animal (“Related Material”), solely for research conducted by the buyer in accordance with all of the following requirements. No right is given to use this Product or Related Material for any other purpose, including, but not limited to, use in drugs, in vitro diagnostic purposes, therapeutics, or in humans. (i) The buyer shall not sell or otherwise transfer Products (including without limitation any material that contains a Licensed Product in whole or part) or any Related Material to any other person or entity, or use Products or any Related Material to perform services for the benefit of any other person or entity, (ii) the buyer shall use only the purchased amount of the Products and components of the Products, and shall use any Related Material, only for its internal research and not for (a) the practice, performance or provision of any method, process or service, or (b) the manufacture, sale, use, distribution, disposition or importing of any product, in each case (a) or (b) for consideration, or on any other commercial basis (“Commercial Purpose”), (iii) the buyer shall use Licensed Products and any Related Material in compliance with all applicable laws and regulations, including without limitation applicable human health and animal welfare laws and regulations, and (iv) the buyer shall indemnify, defend and hold harmless MIT, Harvard and The Broad and their current and former trustees, directors, officers, faculty, affiliated investigators, students, employees, and agents and their respective successors, heirs and assigns (“Indemnitees”), against any liability, damage, loss, or expense (including without limitation reasonable attorneys’ fees and expenses) incurred by or imposed upon any of the Indemnitees in connection with any claims, suits, investigations, actions, demands or judgments arising out of or related to the exercise of any rights granted to the buyer, or any breach of the rights granted hereunder by the buyer.
391 LIMITED USE LABEL LICENSE: RESEARCH USE ONLY Notice to Purchaser: This product is the subject to a license granted to Takara Bio USA, Inc. and its Affiliates from Caribou Biosciences, Inc., and this product is transferred to the end-user purchaser (“Purchaser”) subject to a “Limited Use Label License” conveying to the Purchaser a limited, non-transferable right to use the product, solely as provided to Purchaser, together with (i) progeny or derivatives of the product generated by the Purchaser (including but not limited to cells), and (ii) biological material extracted or derived from the product or its corresponding progeny or derivatives (including but not limited to cells) (collectively, the product, and (i) and (ii) are referred to as (“Material”) only to perform internal research for the sole benefit of the Purchaser. The Purchaser cannot sell or otherwise transfer Material to a third party or otherwise use the Material for any Excluded Use. “Excluded Use” means any and all: (a) commercial activity including, but not limited to, any use in manufacturing (including but not limited to cell line development for purposes of bioproduction), product testing, or quality control; (b) preclinical or clinical testing or other activity directed toward the submission of data to the U.S. Food and Drug Administration, or any other regulatory agency in any country or jurisdiction where the active agent in such studies comprises the Material; (c) use to provide a service, information, or data to a third party; (d) use for human or animal therapeutic, diagnostic, or prophylactic purposes or as a product for therapeutics, diagnostics, or prophylaxis; (e) activity in an agricultural field trial or any activity directed toward the submission of data to the U.S. Department of Agriculture or any other agriculture regulatory agency; (f) high throughput screening drug discovery purposes (i.e., the screening of more than 10,000 experiments per day) as well as scale-up production activities for commercialization; (g) modification of human germline, including editing of human embryo genomes (with the sole exception of editing human embryonic stem (ES) cell lines for research purposes) or reproductive cells; (h) self-editing; and/or (i) stimulation of biased inheritance of a particular gene or trait or set of genes or traits (“gene drive”). It is the Purchaser’s responsibility to use the Material in accordance with all applicable laws and regulations. For information on obtaining additional rights, including commercial rights, please contact licensing@cariboubio.com or Caribou Biosciences, Inc., 2929 7th Street, Suite 105, Berkeley, CA 94710 USA, Attn: Licensing.
396 Sigma-Aldrich CRISPR Use License Agreement This Product and its use are the subject of one or more of the following issued patents and patent applications: Australia Patent Nos. 2013355214; 2017204031; and 2018229489; Canada Patent Nos. 2,891,347 and 2,977,152; China Patent No. CN105142669; European Patent Nos. EP 2 928 496 B1; EP 3 138 910 B1, 3 138 911 B1, EP 3 138 912 B1, EP 3 360 964 B1, EP 3 363 902 B1; Israel Patent No. IL238856; Singapore Patent No. 11201503824S; South Korea Patent Nos. 10-1844123 and 10-2006880; and U.S. Patent Application Serial Nos. 15/188,911; 15/188,924; 15/188,927; 15/188,931; and 15/456,204 (the “Patent Rights”). The purchase of this Product conveys to you (the “Buyer”) the NON-TRANSFERABLE right to use the Product for Licensed Research Use (see definition below) subject to the conditions set out in this License Agreement. 1. “Licensed Research Use” means any use for research purposes, except: (i) Buyer may not sell or otherwise transfer the Product (including without limitation any material that contains the Product in whole or part) or any Related Material to any other third party (except that you may transfer the Product, or any Related Material to a bona fide collaborator or contract research organization), or use the Products or any Related Material to perform services for the benefit of any other third party; (ii) Buyer may use only the purchased amount of the Product and components of the Product, and shall use any Related Material, only for your internal research within the Field, and not for any Commercial Purposes; (iii) Buyer shall use the Product and any Related Material in compliance with all applicable laws and regulations, including without limitation applicable human health and animal welfare laws and regulations; and (iv) the Buyer shall indemnify, defend, and hold harmless SIGMA and their current and former directors, officers, employees and agents, and their respective successors, heirs and assigns (the “Indemnities”) against any liability, damage, loss, or expense (including without limitation reasonable attorneys’ fees and expenses) incurred by or imposed upon any of the Indemnitees in connection with any claims, suits, investigations, actions, demands or judgments arising out of or related to the exercise of any rights granted to the Buyer hereunder or any breach of this License Agreement by such Buyer. 2. For purposes of Section 1 above, the following definitions shall apply: “Commercial Purposes” means (a) the practice, performance or provision of any method, process or service, or (b) the manufacture, sale, use, distribution, disposition or importing of any product, in each case (a) or (b) for consideration, or on any other commercial basis. “Field” means use as a research tool for research purposes; provided, however, that notwithstanding the foregoing, the Field shall expressly exclude (a) any in vivo and ex vivo human or clinical use, including, without limitation, any administration into humans or any diagnostic or prognostic use, (b) the creation of transgenic rodent models and/or derivatives thereof (including, but not limited to, rodents’ cells and rodents’ organs) by for-profit entities, (c) any in vivo veterinary or livestock use, or non-research agricultural use, or (d) use as a testing service, therapeutic or diagnostic for humans or animals. “Related Materials” means any progeny, modification or derivative of a Product. 3. Your right to use the Product will terminate immediately if you fail to comply with these terms and conditions. You shall, upon such termination of your rights, destroy all Product, Related Materials, and components thereof in your control, and notify SIGMA of such in writing. For information on purchasing a license to this Product for purposes other than Licensed Research Use, contact your local SIGMA sales representative, or call +1 800-325-3010.

The Guide-it In Vitro Transcription Kit contains the necessary reagents for the transcription and cleanup of your sgRNA sequences. It is combined with the Guide-it sgRNA Screening Kit as part of the Guide-it Complete sgRNA screening system which can be used for the simple production, cleanup, and evaluation of single guide RNAs (sgRNAs) for CRISPR/Cas9 studies.

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 Image Data

Back

Production of sgRNA by in vitro transcription and testing for cleavage using recombinant Cas9 protein

Production of sgRNA by in vitro transcription and testing for cleavage using recombinant Cas9 protein
Schematic highlighting the key steps for using Guide-it kits for the production of high amounts of single guide RNA (sgRNA) and screening for target site cleavage efficacy using recombinant Cas9 protein.

Back

sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System

sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System
sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System. Panel A. 20 µl of each sgRNA in vitro transcription (IVT) reaction was incubated for 4 hr at 37°C. The yield for each transcribed sgRNA was quantified using a NanoDrop spectrophotometer. Panel B. Each sgRNA IVT reaction was a run on an Agilent Bioanalyzer to assay for sample quality.

Back

sgRNA quality produced via in vitro transcription

sgRNA quality produced via in vitro transcription
sgRNA quality produced via in vitro transcription. sgRNAs produced using the Guide-it sgRNA In Vitro Transcription Kit were compared with sgRNAs produced using a competitor’s kit. The Guide-it kit produced a high-quality, single band for each reaction, while the competitor’s kit showed unwanted byproducts.

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Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs

Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs
Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs.

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)
  • 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 Direct PCR Polymerase Mix (1.25 U/µl)
    • 1.25 ml 2X Terra Direct PCR 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.


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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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  • cDNA synthesis kits
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  • Purified total RNA and mRNA
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  • Most popular polymerases
  • High-yield PCR
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  • In-Fusion seamless cloning
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