RecBCD complex (Exonuclease V) from E.coli is a processive DNA helicase and nuclease that participates in the repair of chromosomal DNA through homologous recombination. The hydrolysis of ssDNA and dsDNA is bi-directional (from both the 3´ and 5´ ends) and processive, producing nucleoside monophosphate. Magnesium ion is required for the exonuclease activity, while calcium, nickel, zinc, and copper inhibits the exonuclease activity and calcium allows double-stranded DNA unwinding (helicase activity) without hydrolysis. RecBCD is expressed and purified from E.coli.
Applications- Removal of contaminating bacterial chromosomal DNA in plasmid, fosmid, cosmid, and BAC clone or vector preparations.- Degradation of linear ssDNA and dsDNA while preserving nicked and supercoiled plasmid DNA. Reagents Supplied & Storage Condition- RecBCD (E.coli): 10 unit/㎕, Store at -20°C.- 10x RecBCD Reaction Buffer : Store at 4°C. - RecBCD in 1X RecBCD Reaction Buffer. Incubate at 37°C.
- 500 mM Potassium Acetate, 200 mM Tris-acetate, 100 mM Magnesium Acetate, 10 mM DTT.
10x Reaction Buffer
Storage Buffer- 50 mM Tris-HCl, 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, 0.1% Triton® X-100 Unit Definition- One unit of RecBCD Nuclease degrades 1 nmol of deoxyribonucleotides in linear dsDNA in 30 minutes at 37°C in 1X Reaction Buffer containing 1 mM ATP. QC Tests
- Activity, exo and endonuclease activity test, SDS-PAGE purity, performance tests.