EZ™ DNA End Repair Module is used to repair the end of fragmented DNA. This kit convert DNA with damaged 5’ protruding or 3’ protruding ends to blunt ended DNA having 5´-phosphates and 3´-hydroxyl. These products are generated by using the 5'→3' polymerase and the 3'→5' exonuclease activities of T4 DNA Polymerase. T4 DNA Polynucleotide Kinase and ATP are also included in the kit for phosphorylation of the 5'-ends of the blunt-ended DNA for subsequent ligation to next-generation sequencing adaptors or other cloning vectors.
DNA repaired by DNA End Repair enzyme mix can be subsequently converted to DNA having 3´ dA-tails with the dA-tailing Module or used directly for blunt end cloning or blunt ended adaptor ligation.
- DNA sample preparation
- End repair of 1-5 μg fragmented DNA
Figure 1. End repair of DNA ends with 3’, 5’ overhangs
Oligoduplex with 3’, 5’ overhangs and non-phosphorylated DNA were repaired using EZ™ DNA END Repair Module. (A) Reaction products were analyzed on 15% native PAGE (B) and on 15% denaturing PAGE.