High Concentration End-Repair Mix
This high-concentration formulation of the End-Repair Mix is compatible with applications requiring >1 microgram of DNA to be prepared for blunt-end ligation. The mix converts DNA containing damaged or incompatible 5′- and/or 3′-protruding ends to 5′-phosphorylated, blunt-ended DNA. The conversion to blunt-ended DNA is accomplished by exploiting the 5′→ 3′ polymerase and 3′→ 5′ exonuclease activities of T4 DNA Polymerase (P7080L). T4 Polynucleotide Kinase (Y9040L) ensures that the ends of the blunt-ended DNA fragments are 5′-phosphorylated for subsequent ligation by T4 DNA Ligase (L6030-HC-L).
Purified from strains of E. coli that express the recombinant T4 DNA Polymerase, and T4 Polynucleotide Kinase genes, respectively.
1 mM Tris-HCl500 mM NaCl100 mM MgCl250 mM DTT0.25% Triton-X 100pH 7.5 @ 25 °C
100 mM KCl10 mM Tris-HCl0.1 mM EDTA1 mM DTT0.1% Triton X-10050% glycerolpH 7.4 at 25 °C
storge at -25 to -15 °C.