This product is an in vitro sgRNA screening kit that can be used to validate the cleavage efficiency of sgRNA (paired with Cas9) in vitro. Using PCR to amplify DNA template containing sgRNA target site, the purified target DNA can be cleaved by sgRNA and Cas9 protein complex, and the cleavage efficiency of Cas9 and sgRNA complex can be detected by agarose gel electrophoresis. This product only contains Cas9 reagents for cleavage experiments, PCR amplification reagents and DNA purification reagents are not included. The entire cleavage experiment can be completed within 2 hours, providing a convenient tool for rapid screening of effective targets.
CRISPR / Cas9 is a breakthrough genome editing technology, with its high efficiency, easy to operate, convenient characteristics, greatly facilitating the gene editing experiments, for biological gene knockout, knock-in, mutation provides a powerful new tool.Cas9 (CRISPR-associated protein 9) is a double-stranded DNA endonuclease, sgRNA ( Cas9 (CRISPR-associated protein 9) is a double-stranded DNA endonuclease, sgRNA (Single Guide RNA) can be combined with Cas9 to form an active ribonucleoprotein (RNP), which guides Cas9 nuclease to reach the DNA target point for cutting, resulting in DNA double-stranded breaks, which may cause gene knockout or knock-in during cellular repair of the damage, and ultimately achieve the purpose of modifying the genomic DNA.
Multiple sgRNAs can be designed according to the target gene, but the efficiency of Cas9 cleavage of DNA mediated by different sgRNAs is inconsistent. Although the non-specific cleavage (off-target) of Cas9 targets can be predicted by bioinformatics when designing the sgRNAs, it is not representative of the actual cleavage efficiency. Therefore, the efficiency of sgRNAs can be verified in vitro before using CRISPR/Cas9 to establish knockout or knock-in organisms, and the selection of sgRNAs that can mediate a higher cutting efficiency can significantly save the overall experimental time, reduce the workload and improve the success rate of gene editing.
Package 2-1 components are as follows (stored at -80°C):
| individual parts making up a compound | norm |
| Control sgRNA (160 ng / μl) *1 | 10 μl |
Package 2-2 is grouped as follows (stored at -20°C):
| individual parts making up a compound | norm |
| 10X Cas9 Digestion Buffer | 150 μl |
| Cas9 Nuclease.,S. pyogenes(160 ng / μl) | 225 μl |
| Control DNA Fragment (50 ng / μl) *2 | 20 μl |
| RNase free water | 1 ml X 2 pcs |
Preservation Temperature:
Package 2-1 -80℃ Storage
Package 2-2 -20℃ Storage
Transportation temperature:
Package 2-1 Dry Ice Transportation
Package 2-2 -20°C Ice Pack Transportation or Dry Ice Transportation
