Restriction enzymes are a class of endonuclease enzymes that recognize specific sequences of deoxyribonucleotides and cut the phosphodiester bond between two deoxyribonucleotides at specific sites in each chain. BsaI is an IIS restriction enzyme that recognizes non-palindromic sequences and cuts sticky ends outside the recognition sequence.SourceE.coliRestriction Enzyme CuttingSite5’…GGTCTC(N)1↓…3’3’…CCAGAG(N)5↑…5’Enzyme Activity DefinitionThe amount of enzyme required to fully digest 1 μg λDNA Dcm-HindIII fragments within 1 h in a 50 μl reaction system at 37℃ is defined as 1 activity unit (U).ProtocolExample 1: DNA rapid enzyme digestion(1) Configure the mixture as shown in the following table:ComponentGenome DNA PCR ProductPlasmid DNARNase-free WaterTo 50 μlTo 30 μlTo 20 μl10× Cut Reaction Buffer5 μl3 μl2 μlDNA10 μl (5 μg)10 μl (0.2 μg)2 μl (1 μg)BsaI5 μl1 μl1 μl(2) Gently mix, do not oscillate the vortex, and then instantaneous centrifugation.(3) Reaction at 37℃ for 15 min (plasmid), or 15-30min (PCR product), or 30-60 min (genomic DNA).(4) Reaction at 80℃ for 20 min can deactivate the enzyme and terminate the reaction (optional).Example 2: It is suitable for extended reaction system of plasmidsRNase-free WaterTo 20 μlTo 20 μlTo 30 μlTo 40 μlTo 50 μl10× Cut Reaction Buffer2μl2 μl3 μl4 μl5 μlPlasmid1 μg2 μg3 μg4 μg5 μgBsaI1 μl2 μl3 μl4 μl5 μlCautions(1) The methylation of Dcm, CpG and EcoB I will impair the cutting activity of this product.(2) Unpurified PCR products have a certain ionic strength. In addition, since DNA polymerase also has exonuclidene activity, it will affect the enzyme digestion products. Therefore, cloning and other operations need to be performed in the following step.(3) This product is for scientific research only and shall not be used for other purposes
Specifictaions: BioReagent, PCR Reagent, for DNA synthesis, Suitable for molecular biology, EnzymoPure™, For In Vitro Transcription, for DNA and RNA applications, 20 U/μl
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- UPC:
- 51281606
- Condition:
- New
- Availability:
- 4-8 weeks
- Weight:
- 1.09 Ounces
- HazmatClass:
- No
- WeightUOM:
- LB
- MPN:
- L359050-1KU
- Product Size:
- 1KU