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LbuCas13a Nuclease
LbuCas13a nuclease is a crRNA-mediated nuclease. By designing single-stranded RNA labeled with fluorescent moieties or other small molecules at both more
Size
100pmol 500pmol 1000pmol
Quantity
Product Description
LbuCas13a nuclease (also known as C2c2) is derived from Leptotrichia buccalis bacterium strain. LbuCas13a belongs to class II type VI CRISPR effector proteins, which are crRNA-mediated nuclease that activate accessory cleavage activity upon recognition and cleavage of target RNA, and can non-specifically cleave single-stranded RNA (ssRNAs) in the system. By designing single-stranded RNA labeled with fluorescent moieties or other small molecules at both ends, the detection and signal amplification of the RNA template by CRISPR/Cas13a can be realized. The results can be observed by fluorometer and test strips.
Product Components
| Component | EDE0002-100 | EDE0002-500 | EDE0002-1000 |
| LbuCas13a Nuclease | 5 μM*20 μL (100pmol) | 5 μM*100 μL (500 pmol) | 5 μM*200 μL (1000 pmol) |
| LbuCas13a Cleavage Buffer (5×) | 500 μL*1 tube | 500 μL*4 tube | 1mL*4 tube |
Test Reaction System
| Component | Final Concentration | Volume ( μL) |
| 5 μM LbuCas13a | 50 nM | 0.25 |
| 5 × Cleavage Buffer | 1× | 5 |
| 25 U RNase Inhibitor | 1 U | 1 |
| 500 nM crRNA | 50 nM | 2.5 |
| 4 μM ssRNA Reporter | 400 nM | 2.5 |
| 1 μM RNA target | 50 nM | 1.25 |
| DEPC H2O | ||
| Total | 25 μL | 25 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 Cas13 collateral cleavage at different concentrations. The horizontal axis is the reaction time, and the vertical axis is the test results of Suzhou Krui MA-1610 constant temperature amplification instrument. As can be seen from the figure, 5×10^9 copies of target RNA can efficiently activate the LbuCas13a enzyme, completely cut the RNA reporter probe within 10 minutes, and reach the fluorescence peak.
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.
