Double-stranded DNA fragments for library construction or large-scale sequencing are often generated by mechanically shearing larger (genomic) DNA, a process that primarily leaves uneven ends. Large DNA may also be cut with restriction enzymes to generate smaller fragments for subcloning. Sheared or restricted DNA fragments must be “end-repaired” before ligation into blunt-end cloning vectors (e.g., Lucigen’s CloneSmart®, CopyRight®, or BigEasy® systems).
The DNATerminator End Repair Kit creates blunt, 5' phosphorylated ends on any type of sheared or restriction-digested DNA fragment, ensuring the highest possible cloning efficiency (Figure 1).
Figure 1. Effect of DNA shearing and end-repair methods on library construction efficiency. Shotgun libraries were constructed using 250 ng of lambda DNA sheared by sonication (Son) or by the GeneMachines® HydroShear™ device (HS). Sheared DNA was repaired with T4 DNA polymerase and Klenow fragment (TK) or DNATerminator End Repair Kit (DT) and cloned using Lucigen’s CloneSmart® system. Values are colony forming units (cfu) per library.
|Citations||DNATerminator® End Repair Kit - Citations||LINK|
|SDS||SDS073 40035-1_40035-2_40035-3 DNATerminator End Repair Kit.pdf|