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LbCas12a Nuclease
LbCas12a nuclease is a crRNA-mediated nuclease that recognizes and cleaves double-stranded DNA (dsDNA) in the presence of a PAM (Protospacer Adjacent more
Size
100pmol 1000pmol 2000pmol
Quantity
Product Description
LbCas12a nuclease (also known as Cpf) is derived from Lachnospiraceae bacterium ND2006 strain. LbCas12a belongs to class 2 type V CRISPR effector proteins, and is a crRNA-mediated nuclease that recognizes and cleaves double-stranded DNA (dsDNA) in the presence of a PAM (Protospacer Adjacent Motif) sequence (TTN). For single-stranded DNA (ssDNA) cleavage, the PAM sequence is not required. Upon binding to complementary ssDNA or dsDNA, the trans-cleavage activity of LbCas12a on non-specific ssDNA is activated. By designing single-stranded Reporter DNA labeled with fluorescent moieties or other small molecules at both ends, the detection and signal amplification of the DNA template by Cas12a can be realized. The results can be observed by fluorometer and test strips.
Product Components
| Component | EDE0005-100 | EDE0005-1000 | EDE0005-2000 |
| LbCas12a Nuclease | 1 μM*100 μL (100pmol) | 1 μM*500 μL*2 tube (1000 pmol) | 1 μM*500 μL*4 tube (2000 pmol) |
| LbCas12a Cleavage Buffer (10×) | 500 μL*1 tube | 1mL*4 tube | 1mL*7 tube |
Test Reaction System
| Component | Final Concentration | Volume ( μL) |
| 10× Cleavage Buffer | 1× | 3 |
| 1 μM LbCas12a Nuclease | 33 nM | 1 |
| 500 nM crRNA | 33 nM | 2 |
| 2 μM ssDNA Reporter | 400 nM | 6 |
| 1 μM DNA target | 3.5 nM | 0.1 |
| DEPC H2O | ||
| Total | 30 μL | 30 μL |
Reaction Conditions
Use a real-time fluorescence quantitative PCR machine or an isothermal amplification instrument to detect the fluorescence signal. The reaction should occur at 37°C, and the fluorescence signal should be collected every 30 seconds. Alternatively, the fluorescence signal can be directly observed under a UV light.
Cross-Analysis of Competitive Performance
Fig 1. Results of Cas12a collateral cleavage at different company product. The abscissa is the reaction time, and the ordinate is the detection result of Bio-rad CFX96 fluorescence PCR instrument. As can be seen from the figure, under the same conditions, the cutting efficiency of this product is significantly higher than that of similar products.
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.
