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SpCas9 Nuclease
SpCas9 Nuclease is an sgRNA-guided DNA endonuclease. In the presence of a PAM (proto-spacer adjacent motif) sequence in the target DNA, Cas9 Nuclease more
Size
100pmol 1000pmol
Quantity
Product Description
Cas9 Nuclease (SpCas9), derived from the S. pyogenes strain, is an sgRNA-guided DNA endonuclease that cleaves double-stranded DNA. In the presence of a PAM (proto-spacer adjacent motif) sequence in the target DNA, Cas9 Nuclease can specifically cut the target double-stranded DNA under the guidance of sgRNA, creating blunt-end double-strand breaks. The PAM sequence is essential for Cas9 to recognize and cleave the target DNA, with the cleavage site located within the target sequence, 3 base pairs away from the PAM region. Cas9 Nuclease can also be applied in experiments such as in vitro target DNA cleavage and cloning of specific fragments.
Product Components
| Component | EDE0008-100 | EDE0008-1000 |
| SpCas9 Nuclease | 1 μM*100 μL (100 pmol) | 1 μM*1mL (1000 pmol) |
| Buffer (5x) | 100 μL*1 tube | 500 μL*1 tube |
Test Reaction System
| Component | Volume (μL) | Final Concentration |
| 5× Cleavage Buffer | 4 | 1x |
| SpCas9 Nuclease (1 μM) | 5 | 250 nM |
| 1000 nM sgRNA | 0.5 | 250 nM |
| 1 μM DNA | 0.5 | 25 nM |
| DEPC H2O | Add to 20 μL | - |
Reaction Conditions
Incubate at 37°C for 30 minutes. The results can be analyzed via non-denaturing nucleic acid gel electrophoresis.
Storage Conditions and Shelf Life
The product is stable for 1 year when stored at -20°C. For long-term storage, it is recommended to store at -80°C. It is advised to aliquot the product based on the frequency of use to avoid repeated freeze-thaw cycles.
