V32101T SpCas9 Powder (Single Reaction)

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V32101T SpCas9 Powder (Single Reaction)

  • Specifications:
    • 48 T
    • 96 T

Article number:


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Product overview


SpCas9 is a DNA endonuclease guided by crRNA and tracrRNA (or sgRNA alone), which is derived from S. pyogenes. SpCas9 specifically recognizes and cleaves dsDNA target in a PAM (NGG)-dependent manner. The cleavage site of Cas9 in the target sequence is 3 bps away from the PAM site. In addition, SpCas9 can also specifically cleave ssDNA or ssRNA in the presence of DNA PAMmer sequences (for details, please refer to PMID: 24476820; PMID: 25274302). SpCas9 can also be used in in vitro experiments for cleaving target DNA and cloning target fragments (for details, please refer to PMID: 26323354).

This product is lyophilized glycerol-free SpCas9 nuclease, which can be shipped and stored at room temperature. After being dissolved with the Reconstitution Solution, it shows the same performance as the non-lyophilized type of SpCas12b Nuclease (#32101, ToloBio). This product can be provided in a variety of lyophilized forms such as lyophilized powder and lyophilized beads to meet the needs of different downstream applications.




This product is stored and shipped at room temperature. After reconstitution, the liquid enzyme should be stored at -20 ℃ or below and is stable for at least one month. Repeated freezing and thawing should be avoided for reconstituted liquid enzyme.




SpCas9 Powder(Single Reaction)




SpCas9 Lyophilized Powder (8-TUBE STRIPS)

8 T 48 T 96 T

10 × HOLMES Buffer for Cas9

1 mL 1 mL 1 mL

· The amount of SpCas9 for a single reaction is 5 pmol and is recommended for a 20 μL CRISPR reaction system.



Experiment Procedure

SpCas9 (Cat# V32101T/V32101S) CRISPR Cis-cleavage Experiment

1.Prepare the reaction mixture of CRISPR Cis-cleavage Experiment on ice as below.



Final concentration 

10 × HOLMES Buffer for Cas9(#32041)

2 μL 1 ×

10 μM sgRNA

0.5 μL 250 nM

1 μM Target dsDNA

0.5 μL 25 nM

Nuclease-free water

Up to 20 μL  

2.Dissolve the lyophilized powder/bead for a single reaction using the 20 μL reaction mixture prepared above.

3.Put the reaction mixture at 37℃for 30 min and then inactive the enzyme at 85℃for 5 min . Analyze products using nucleic acid electrophoresis.



Experimental Results


Product Instruction

#V32101 SpCas9 Nuclease (Lyophilized)




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