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  more

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100pmol 500pmol 1000pmol

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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 EDE0006-500 EDE0006-1000
AapCas12b Nuclease 5 μM*10 μL (50pmol) 5 μM*100 μL (500 pmol) 5 μM*200 μL (1000 pmol)
AapCas12b Cleavage Buffer (10×) 40 μL*1 tube 400 μL*1 tube 800 μL*1 tube

Test Reaction System

Component Final Concentration Volume ( μL)
10× Cleavage Buffer 2 μL
5 μM AapCas12b Nuclease 150-250 nM 0.8 μL(200 nM)
500 nM crRNA 150-250 nM 8 μL(200 nM)
4 μM 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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