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.

Reaction Condition
- RecBCD in 1X RecBCD Reaction Buffer. Incubate at 37°C.

10x Reaction Buffer
- 500 mM Potassium Acetate, 200 mM Tris-acetate, 100 mM Magnesium Acetate, 10 mM DTT.

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.