Source: Recombinant Cas12b, produced through genetic engineering, expressed in Escherichia coli.
一、 Product Introduction
Crispr Cas12b nuclease (also known as C2c1) is an RNA mediated endonuclease that can specifically cleave target double stranded DNA in the presence of PAM. The Cas12b protein is smaller than Cas9 and Cas12a, and originates from Alicyclobacillus acidoterrestris, an acidophilic and heat-resistant bacterium with higher cutting activity. The optimal cleavage reaction temperature for Cas12b is 48 ℃. Similar to the same family of Cas12a, Cas12b also recognizes the PAM sequence of 5 ‘- TTN and produces sticky ends after DNA cleavage; However, Cas12b is dual RNA directed and relies on crRNA and tracrRNA (or sgRNA formed after connection).
In addition, Cas12b can not only be used for target dsDNA cleavage, but also for rapid detection of target nucleic acids, as detailed in the HOLMESv2 nucleic acid rapid detection technology. This product (including Cas12b enzyme) does not contain any other nuclease activity except for Cas12b. Cas12b is a nuclease based on the CRISPR system. Compared to Cas9, Cas12b has high specificity and efficiency, and can be used to knock out and/or insert genes in cells. The Cas12b system was developed and modified by the team led by Zhang Feng.
二、Product features
1 | Appearance | transparent liquid |
2 | Purity | ≥90%(SDS-PAGE/SEC-HPLC) |
3 | The residual amount of host protein | <100ng/mg |
4 | Bacterial endotoxins | <10EU/mg |
5 | The residual amount of host DNA | <10ng/mg |
6 | DNA enzyme residue | negative |
7 | RNA enzyme residue | negative |
8 | Exonuclease activity | ≥80% |
9 | Concentration | ≥5mg(ABS) |
三、Product application
When Cas12b recognizes and cleaves target double stranded DNA under the guidance of guide RNA, its “accessory cleavage” activity is activated, which can efficiently cleave non-specific single stranded DNA (ssDNA) in the system. By designing ssDNA probes labeled with fluorescent groups or other markers at both ends, CRISPR/Cas12b can detect DNA templates and amplify signals, thereby achieving target detection. This system has high sensitivity and strong specificity.
The commonly used detection methods include real-time fluorescence and colloidal gold chromatography. At present, the system has been widely used in gene editing of microorganisms, plants, and animals, and has great practical application value in molecular diagnosis.
Used for direct detection of DNA in molecular diagnosis, if RNA detection requires binding to enzymes that transcribe DNA into RNA.
四、Storage and transportation stability
1、Storage stability: valid for two years at temperatures ranging from -25 to -15 ℃.
2、Transportation stability: refrigerated transportation, stable activity.
3、Attention: It is not recommended to store this product at -80 ℃. Repeated freezing and thawing will reduce the enzyme activity of this product.
五、Product advantages
1. No animal origin: Recombinant production, no exogenous virus contamination, and no animal source raw materials are used in the production process.
2. Stable quality: Batch production (stable production supply of 100g) can ensure stable and continuous batch production; There is no difference between product batches, and the quality is stable.
3. High purity: high specific activity; The residual host protein is less than the limit requirement for biological products.
4. Compliance with regulatory requirements: Production equipment and environment comply with relevant regulatory requirements, and the production process fully complies with GMP guidance principles.
5. Complete quality documents: According to customer requirements, relevant regulatory support documents can be provided.
六、Related products
Recombinant sumo enzyme;
Recombinant TEV enzyme;
Recombinant intestinal kinase.
For scientific research and production purposes only, not for human use.