Description
CRISPR/Cas9 is a technique in which RNA directs Cas nucleases to perform specific DNA modifications on targeted genes, resulting in an acquired immune defense mechanism evolved by bacteria and archaea in response to attacks by bacteriophages and exogenous plasmids. In modern biotechnology research, this system guides the nuclease Cas9 protein to cut double-stranded DNA at the target site paired with sgRNA by artificially optimized sgRNA (Single Guide RNA), causing DNA breaks. Due to the repair of DNA by non-homologous terminal repair mechanism or homologous recombination mechanism in organisms, gene shifting, mutation, replacement or deletion occur, resulting in loss of gene function.
The Hifair™ Precision sgRNA Synthesis Kit uses a T7 RNA polymerase to efficiently transcribe spCas9 sgRNA. This kit contains a specific sequence that is recognized by the Cas9 protein and acts as a scaffold, and part of this sequence can partially overlap with the user-designed specific sequence of interest. After annealing, complementary strands are formed, which are filled by DNA polymerase. The resulting dsDNA template for transcription is generated. Using the reagents provided and user-designed specific DNA sequences, this kit can obtain 20-100 μg of functional sgRNA in 4 hours in a single tube reaction.
Components
|
No. |
Name |
11355ES25 (25 T) |
11355ES50 (50 T) |
|
11355-A |
10×sgRNA Enzyme Mix |
50 μL |
100 μL |
|
11355-B |
10×sgRNA Reaction Buffer |
50 μL |
100 μL |
|
11355-C |
2×Canace Enzyme Mix |
312.5 μL |
625 μL |
|
11355-D |
Scaffold Template |
31.25 μL |
62.5 μL |
|
11355-E |
NTPs（25mM each） |
200 μL |
400 μL |
|
11355-F |
Control sgRNA Oligo（10μM） |
10 μL |
20 μL |
Shipping and Storage
-25℃ ~ -15℃ storage, valid for 2 years.