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AapCas12b Nuclease
AapCas12b nuclease is a crRNA-mediated nuclease, with an optimal reaction temperature of 60°C, making it more heat-resistant compared to Cas13a and Cas12a. This property makes it highly suitable for field nucleic acid detection applications when combined with loop-mediated isothermal amplification (LAMP). 更多
货号 : EDE0006
Size
100pmol 500pmol 1000pmol
数量
Product Description
AapCas12b nuclease (also known as C2c1) is derived from Alicyclobacillus acidophilus. It 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 AapCas12b on non-specific ssDNA is activated. AapCas12b has an optimal reaction temperature of 60°C, making it more heat-resistant compared to Cas13a and Cas12a. This property makes it highly suitable for field nucleic acid detection applications when combined with loop-mediated isothermal amplification (LAMP).
Product Components
| Component | EDE0006-50 | EDE0006-100 | EDE0006-500 |
| AapCas12b Nuclease | 5 μM*10 μL （50pmol） | 5 μM*20 μL （100pmol） | 5 μM*100 μL （500 pmol） |
| AapCas12b Cleavage Buffer（10×） | 40 μL*1tube | 80 μL*1tube | 400 μL*1tube |
Test Reaction System
| Component | Final Concentration | Volume ( μL) |
| 10× Cleavage Buffer | 1× | 2 μL |
| 5 μM AapCas12b Nuclease | 150-250 nM | 0.8 μL（200 nM） |
| 500 nM crRNA | 150-250 nM | 8 μL（200 nM） |
| 250 nM ssDNA Reporter | 150-250 nM | 1 μL（200 nM） |
| 1 μM DNA target | 150-250 nM | 4 μL（200 nM） |
| DEPC H2O | ||
| Total | 20 μL | 20 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 60°C, and the fluorescence signal should be collected every 30 seconds. Alternatively, the fluorescence signal can be directly observed under a UV light.
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.
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