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V32118E AapCas12b Powder

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V32118E AapCas12b Powder

  • Specifications:
    • 100 µg
    • 10×100 µg

Article number:

#V32118E


Price:Contact our distributors


隐藏域元素占位

Product overview

Description

AapCas12b (previously known as C2c1) is a DNA endonuclease mediated by tracrRNA and crRNA (or sgRNA), which specifically cleaves the dsDNA target in a PAM (TTN)-dependent manner, generating DSBs with sticky ends, and cleave ssDNA target in a PAM-independent manner. In addition, both dsDNA and ssDNA targets can trigger the trans-cleavage activity of AapCas12b, that is, when the AapCas12b binds sgRNA and target DNA to form a ternary complex, the trans-cleavage activity of AapCas12b will be released to non-specifically cleave ssDNA sequences in the reaction system. AapCas12b is derived from Alicyclobacillus acidophilus and produced from E. coli BL21(DE3), with the optimal reaction temperature of 60 °C for in vitro analysis. Therefore, AapCas12b is more suitable for the creature of a “One-Step” CRISPR-Cas detection system when combined with the LAMP isothermal amplification technology.

This product is lyophilized glycerol-free AapCas12b nuclease, which can be shipped and stored at room temperature. After being dissolved with its reconstitution buffer, it shows the same performance as the non-lyophilized type of AapCas12b (C2c1) Nuclease (#32118, ToloBio). This product can be provided in a variety of lyophilized forms such as lyophilized powder and lyophilized beads to meet the needs of different downstream applications

 

 

Storage Conditions

This product is stored and shipped at room temperature. After reconstitution, the liquid enzyme should be stored at -20 °C or below and is stable for at least one month. Repeated freezing and thawing should be avoided for reconstituted liquid enzyme

 

 

Components

AapCas12b Powder

V32118E-SP 

V32118E-01 

V32118E-02 

AapCas12b Lyophilized Powder (SCREW CAP TUBES)

10 μg 100 μg 100 μg × 5

Reconstitution Solution

200 μL 200 μL 1 mL

10 × HOLMES Buffer for Cas12b

1 mL 1 mL 1 mL × 5

· 100 μg of AapCas12b nuclease = 750 pmol. It is recommended to use 150 μL of reconstitution solution to reconstitute 100 μg AapCas12b lyophilized powder, generating 5 μM (5 pmol/μL) liquid enzyme. Once dissolved, the liquid enzyme should be stored at -20 °C or below.

 

 

Experiment Procedure

AapCas12b (Cat # V32118E) CRISPR Trans-cleavage Experiment

1.Prepare the reaction mixture of CRISPR Trans-cleavage Experiment on ice as below.

Components

Volume 

Final concentration 

10 × HOLMES Buffer for Cas12b (#32043)

2 μL

1 × 

5 μM AapCas12b(C2c1)Nuclease

1 μL

250 nM
10 μM sgRNA

0.5 μL

250 nM
1 μM Target DNA  0.5 ~ 5 μL

25 ~ 250 nM

10 μM HOLMES ssDNA Reporter(#31101)  0.5 μL

250 nM

Nuclease-free Water

Up to 20 μL

 

2.Put the reaction mixture in the real-time PCR instrument to detect the fluorescent signal at 60 °C, and collectsthe fluorescent signals every 30 sec.

 

 

Experimental Results

FAQ

Product Instruction

#V32118 AapCas12b (C2c1) Nuclease (Lyophilized)

pdf

875.2KB

References

COA Query

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