Description

LbCas12a (previously known as Cpf1) is a crRNA-guided DNA endonuclease derived from the Lachnospiraceae bacterium ND2006 strain. LbCas12a specifically cleaves the dsDNA target in a PAM (TTN)-dependent manner, generating DSBs with sticky ends, and cleaves ssDNA target in a PAM-independent manner. In addition, both dsDNA and ssDNA targets can trigger the trans-cleavage activity of LbCas12a, that is, when the LbCas12a binds crRNA and target DNA to form a ternary complex,the trans-cleavage activity of Cas12a will be released to non-specifically cleave ssDNA sequences in the reaction system.

This product is lyophilized glycerol-free LbCas12a nuclease, which can be shipped and stored at room temperature. After being dissolved with its reconstitution buffer, it shows the same performance as the non-lyophilized type of LbCas12a (Cpf1) Nuclease (#32108, ToloBio). This product can be provided in a variety of lyophilized forms such as powder and beads to meet the needs of different downstream applications

Components

Storage Conditions

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

Experiment Procedure

LbCas12a (Cat# V32108T/V32108S) CRISPR Trans-cleavage Experiment

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

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

3.Put the reaction mixture in the real-time PCR instrument to detect the fluorescent signals at 37 °C, and collect the fluorescent signals every 30 sec.

Experimental Results