The BeyoCRISPR™ sgRNA Synthesis Kit provides a simple and quick method for in vitro synthesis of large amounts of sgRNA (single guide RNA), using the supplied reagents in the kit and target-specific DNA oligos designed by users. We provide two types of sgRNA synthesis kits: BeyoCRISPR™ One-Step sgRNA Synthesis Kit (D7081) and BeyoCRISPR™ Two-Step sgRNA Synthesis Kit (D7083). The sgRNA yield of the two-step method is twice that of the one-step method.
BeyoCRISPR™ One-Step sgRNA Synthesis Kit (D7081)
BeyoCRISPR™ One-Step sgRNA Synthesis Kit provides all reagents required for sgRNA synthesis except Target-specific DNA Oligo. Cas9 Scaffold Oligo included in this kit is a 80nt oligonucleotide. The users need to design a Target-specific DNA oligo with a 3’-end complementary to the 3’-end of the Cas9 Scaffold Oligo, by which a dsDNA can be synthesized using the supplied Enzyme Mix in the kit. The dsDNA is then used as a template to synthesize the sgRNA by RNA polymerase included in the Enzyme Mix. Please see Figure 1 for the schematic diagram of the one-step sgRNA synthesis method.
Figure 1. Schematic diagram of sgRNA synthesis with the BeyoCRISPR™ One-Step sgRNA Synthesis Kit (Beyotime, #D7081).
Features:
Simple and Efficient: Synthesis of 5-25μg sgRNA in a 20μl reaction within 0.5-4 hours.
Cost-effective: High-quality product at low cost.
Wide Applications: The obtained sgRNA can be mixed with Cas9 nuclease to evaluate the sgRNA-guided cleavage of DNA substrate in vitro, or transfected into cells expressing Cas9 to edit target genes in vivo.
Typical Figures:
Figure 2. Non-denaturing PAGE analysis of sgRNA synthesized by the BeyoCRISPR™ One-Step sgRNA Synthesis Kit (Beyotime, #D7081). Reactions were assembled as described in the product manual. The in vitro transcription reactions were incubated for different time courses as indicated. The synthesized sgRNA with or without DNase I were separated on 15% non-denaturing PAGE. The gel was stained with NA-Red (D0130) at room temperature for 15 minutes, followed by examination by a gel imaging system. The result demonstrates that sgRNA yield is higher with longer duration of in vitro transcription. This figure is for reference only.
Figure 3. Agarose gel electrophoresis of DNA substrate with or without incubation with sgRNA synthesized by the BeyoCRISPR™ One-Step sgRNA Synthesis Kit (Beytime, #D7081). The sgRNA-guided cleavage of DNA substrate was performed as described in the product manual. Lane 1: DNA substrate (2773bp) without sgRNA incubation; Lane 2: DNA substrate (2773bp) incubated with sgRNA was completely cut into two fragments of 2206bp and 567bp. This figure is for reference only. The results may vary depending on different experimental conditions.
BeyoCRISPR™ Two-Step sgRNA Synthesis Kit (D7083)
BeyoCRISPR™ Two-Step sgRNA Synthesis Kit provides all reagents required for sgRNA synthesis except Target-specific DNA Oligo. Scaffold Template provided in this kit is a 80 nt oligonucleotide including the 5’-end overlapping region with the 3’-end of the Target-specific DNA oligos designed by users and the 3’-end sequence complementary to the reverse primer used for the amplification of DNA template. With the reagents included in this kit and the Target-specific DNA Oligo, it takes 0.5h-4h to obtain 10-40μg sgRNA in a 20μl transcription reaction. Please see Figure 4 for the schematic diagram of the two-step sgRNA synthesis method.
Figure 4. Schematic diagram of sgRNA synthesis with the BeyoCRISPR™ Two-Step sgRNA Synthesis Kit (Beyotime, #D7083).
Features:
Simple and Efficient: Synthesis of 10-40μg sgRNA in a 20μl reaction within 0.5 -4 hours. Low Cost: High-quality product at low cost. Wide Applications: The obtained sgRNA can be mixed with Cas9 nuclease to evaluate the sgRNA-guided cleavage of DNA substrate in vitro, or transfected into cells expressing Cas9 to edit target genes in vivo.
Typical Figures:
Figure 5. Non-denaturing PAGE analysis of sgRNA synthesized by the BeyoCRISPR™ Two-Step sgRNA Synthesis Kit (Beyotime, #D7083). Reactions were assembled as described in the product manual. The in vitro transcription reactions were incubated for different time courses as indicated. The synthesized sgRNA with or without DNase I were separated on 15% non-denaturing PAGE. The gel was stained with NA-Red (D0130) at room temperature for 15 minutes, followed by examination by a gel imaging system. The result demonstrates that sgRNA yield is higher with longer duration of in vitro transcription. This figure is for reference only.
Figure 6. Agarose gel electrophoresis of DNA substrate with or without incubation with sgRNA synthesized by the BeyoCRISPR™ Two-Step sgRNA Synthesis Kit (Beytime, #D7083). The sgRNA-guided cleavage of DNA substrate was performed as described in the product manual. Lane 1: DNA substrate(2773bp) without sgRNA incubation; Lane 2: DNA substrate (2773bp) incubated with sgRNA was completely cut into two fragments of 2206bp and 567bp. This figure is for reference only. The results may vary due to different experimental conditions.
Product Information：
| Catalog No. | Product Name | Size | Price |
| D7081S | BeyoCRISPR™ One-Step sgRNA Synthesis Kit | 20T | |
| D7081M | BeyoCRISPR™ One-Step sgRNA Synthesis Kit | 100T | |
| D7083S | BeyoCRISPR™ Two-Step sgRNA Synthesis Kit | 20T | |
| D7083M | BeyoCRISPR™ Two-Step sgRNA Synthesis Kit | 100T | |
| D0511S | Cas9 Nuclease (SpCas9) | 50pmol | |
| D0511M | Cas9 Nuclease (SpCas9) | 250pmol | |
| D0511L | Cas9 Nuclease (SpCas9) | 1000pmol |