Description |
GenCRISPR™ LbCas12a Nuclease is an RNA-guided DNA endonuclease fromLachnospiraceae bacterium. Cas12a (previously known as Cpf1) belongs to the Class 2 Type V CRISPR/Cas system. Different from CRISPR/Cas9 system, the ribonucleoprotein (RNP) complex of CRISPR/Cas12a system is formed by Cas12a and crRNA, it recognizes a T-rich protospacer adjacent motif (PAM) and results in a staggered DNA double-strand break (DSB). After the specific cleavage, Cas12a can also activate collateral cleavage activity towards adjacent non-specific ssDNA sequences. Hence, Cas12a nuclease is a good alternative for Cas9 in certain target DNA editing, and provides a novel strategy for DNA detection. |
crRNA-dependent DNA cleavage
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Source |
Recombinant Cas12a with a C-terminal NLS expressed byE.coli
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Species |
Lachnospiraceae bacterium
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Tag |
C-terminal 6× His Tag
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Apparent Molecular Weight |
146 kDa, on SDS-PAGE under non-reducing conditions
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Concentration |
4 mg/ml
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Storage Buffer |
10 mM Tris-HCl, 300 mM NaCl, 0.5 mM DTT, 50% glycerol, pH 7.4.
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Storage & Stability |
Store at -20 °C for up to 12 months from the date of manufacture. Avoid repeated freeze-thaw cycles.Do not store below -20 °C!
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Accession# |
6KL9_A
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Appearance |
Clear, colorless liquid
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Purity |
≥ 90% by SDS-PAGE
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Concentration by A280 |
4(± 10%) mg/ml
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Bioactivity (in vitro) |
≥ 90%
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Residual DNase |
Undetectable
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Residual RNase |
Undetectable
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Endotoxin Level |
< 0.1 EU/μg
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Figure 1: In vitro digestion efficiency analysis by agarose gel electrophoresis.A 20 μl reaction in 1 × Cas12a Nuclease Reaction Buffer containing 60 ng linearized plasmid, 10 ng crRNA, and 100 ng GenCRISPR™ LbCas12a Nuclease (Cat. No. Z03753) for 30 min at 37°C results in a digestion efficiency of linearized plasmid higher than 90%, as determined by agarose gel electrophoresis.
References |
1. Zetsche, Bernd, et al. "Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system." Cell 163.3 (2015): 759-771. 2.Ledford, Heidi. "Alternative CRISPR system could improve genome editing."Nature News526.7571 (2015): 17. 3.
Chen, Janice S.,et al."CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity."Science360.6387 (2018): 436-439.
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For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.