- Detail
- Parameters
Product name
Magigen CRISPR Cas12 protein | AapCas12b Protein Kit
Magigen CRISPR Cas12 protein product AapCas12b is an RNA-guided endonuclease. It can specifically cleave target double-stranded DNA in the presence of PAM on target DNA.
Catalog
C006S/M
Intended use
This kit is suitablefor the development of detection products based on CRISPRplatform.
Size
100pmol/1000pmol
Contents of the kit
Item
C006S
C006M
10×Cas12b Reaction Buffer
1ml×1Tube
1ml×6 Tube
AapCas12b Protein
50µl（2µM）
100µl（10µM）
1×Cas12b Dilution Buffer
\
1ml×2 Tube
Detection principle
Magigen AapCas12b protein, derived from Alicyclobacillus acidiphilus, is an RNA-mediated endonuclease that binds to and cleaves specific sites of target DNA under the guidance of single-stranded RNA. While Cas12b recognizes and cleaves target double-stranded DNA under the guidance of guide RNA, its collateral cleavage activity is activated, which can efficiently cleave non-specific single-stranded DNA (ssDNA). CRISPR/Cas12b can be used to detect DNA templates and amplify signals by designing ssDNA probes labeled with fluorescent groups or other markers at both ends. Common detection methods include real-time fluorescence and colloidal gold chromatography. At present, this system has been widely used in gene editing of micro organisms, plants and animals, and has great practical application value in molecular diagnosis.
Storage conditions and Expiration
Storage conditions: -20℃, avoid light.
Expiration: 12 months. Avoid repeated freezing-thawing after opening.
Applicable instrument
Microplate Reader, like BioTek Synergy et al. fluorescence quantitative PCR instruments with FAM channel.
Operation procedure
1. Preparation of reaction substrate
The reaction substrate could be nucleic acid amplification products of LAMP / PCR / RPA.
2. Methods
2.1 Recommended reaction system
Component
Reaction volume / final concentration
10×Cas12b Buffer
5 μL
fluorescent probe
40 ~ 160nM
gRNA
20 - 50 nM
AapCas12b Protein
100 - 500nM
template
2 μL
H2O
up to 50 μL
2.2 Set up the program in advance and preheat while using microplate reader.
Filter Set: Green（Ex:485/20, Em: 528/20），Reaction temperature: 45 ~ 65℃, get the fluorescence values every 1 ~ 4 min, time: 30min ~ 1h.
3. 【Interpretation of the results】
CRISPR-Cas negative result and positive result
Product Performances
According to tested results below, using different concentrations of Cas12a protein to cut ssDNA, it can be found that a sufficient amount of Cas12a can cut DNA in about 12 minutes.
Magigen crispr cas12b protein is a qualified cas protein with good price.
The crispr cas12b price will be subject to change without prior notice due to currency fluctuation. All Pictures shown are for illustration purpose only and shall not form part of any offer, contract.
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