T4 UvsX Recombinase

The T4 UvsX Recombinase derived from T4 Phage is the homologue of RecA/Rad51 family. It plays an important role in the repair of double-stranded DNA breaks and in the restarting of replication forks. T4 UvsX Recombinases can be anchored on a single-stranded DNA together with other recombinases and activate them to search for homologous sequences on other double-stranded DNA to further complete the chain replacement.

Storage Temperature

Store at -20 ℃, valid for one year (Avoid repeated freeze-thaw cycles).

Protein Concentration

The concentration of T4 UvsX Recombinase is 2 mg/mL±5% as determined by UV absorption at 280 nm.

Storage Buffer

20 mM Tris-HCl, 0.5 M NaCl, pH 8.0, 50% glycerol.

Heat Inactivation

60℃ for 10 min.

Quality Control

Exonuclease Activity: 2 μg of T4 UvsX Recombinase with 1 μg λ-Hind III digest DNA at 37℃ for 16 hours yields no degradation as determined by agarose gel electrophoresis.

Endonuclease Activity: 2 μg of T4 UvsX Recombinase with 1 μg λDNA at 37℃ for 16 hours yields no degradation as determined by agarose gel electrophoresis.

Nicking Activity: 2 μg of T4 UvsX Recombinase with 1 μg pBR322 at 37℃ for 16 hours yields no degradation as determined by agarose gel electrophoresis.

RNase Activity: 2 μg of T4 UvsX Recombinase with 1.6 μg MS2 RNA for 16 hours at 37℃ yields no degradation as determined by agarose gel electrophoresis.

E. coli DNA: 2 μg of T4 UvsX Recombinase is screened for the presence of E. coli genomic DNA using TaqMan qPCR with primers specific for the E. coli 16S rRNA locus. The E. coli genomic DNA contamination is ≤1 E. coli genome.

Reaction Condition

1× UvsX Recombinase Buffer： 20 mM Tris-acetate, pH 7.8 100 mM Potassium acetate 10 mM Magnesium acetate 1 mM DTT

1. DNA isothermal amplification
2. RPA amplification