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Cat. No.: ENLbCas12a-100 (for 100pmol)

Cat. No.: ENLbCas12a-1k (for 1000pmol)

Description

Enhanced LbCas12a is a high-activity mutant of the wild-type LbCas12a nuclease, obtained through rational engineering and optimization. Like the wild-type enzyme, enhanced LbCas12a is guided by crRNA to specifically recognize double-stranded DNA (dsDNA) targets containing a PAM sequence (5′-TTN-3′), introducing double-strand breaks with sticky ends.

In addition, enhanced LbCas12a can recognize and cleave single-stranded DNA (ssDNA) targets in a PAM-independent manner. Both dsDNA and ssDNA targets can activate the enzyme’s collateral (trans) cleavage activity. Once enhanced LbCas12a forms a ternary complex with crRNA and the target DNA, it is activated to indiscriminately degrade non-specific ssDNA molecules present in the reaction system.

Therefore, enhanced LbCas12a can be applied not only for in vitro site-specific cleavage of dsDNA but also for rapid nucleic acid detection assays.

Components

• Enhanced LbCas12a
• 10× Buffer for Cas12a  

Storage Conditions

• Store all components at –20 °C.
• Transport at ≤0 °C.

Notes

• The direct repeat (DR) sequence of crRNA for enhanced LbCas12a is identical to that of wild-type LbCas12a. Therefore, enhanced LbCas12a can be directly substituted for wild-type LbCas12a in reaction systems.
• Cas12a nucleases from different species may vary slightly in their PAM requirements. Most Cas12a enzymes recognize either TTN or TTTN PAM sequences.
• Cis-cleavage activity: Guided by crRNA, Cas12a specifically cleaves target DNA. dsDNA targets require a PAM site, whereas ssDNA targets are cleaved in a PAM-independent manner.
• Trans-cleavage activity: In the presence of target DNA, Cas12a forms a ternary complex with crRNA and the target DNA (Cas12a/crRNA/target DNA). Once activated, Cas12a exhibits trans-cleavage activity, indiscriminately degrading non-specific ssDNA molecules within the reaction system.
• To prevent RNase contamination, maintain a clean and tidy work area. Always wear clean gloves and a mask during operation, and ensure that all consumables (pipette tips, centrifuge tubes, etc.) are RNase-free.
• Cas12a protein is heat-sensitive and prone to inactivation. Reactions should be assembled on ice, and the enzyme should be returned to –20 °C immediately after use.
• For research use only.