In the paper, according to RSAS, "Modrich demonstrated the requirement of DNA polymerase III, exo-nuclease I, and DNA ligase
for mismatch repair.
The recent observations that DNA ligase
III levels are decreased in the absence of ATM may also explain these results (Sharma et al.
1997) Mammalian DNA double-strand break repair protein XRCC4 interacts with DNA ligase
Then, the purified PCR product was ligated to pMD18-T vector (an T cloning vector) by T4 DNA ligase
at 16 oC overnight.
05 [micro]l (40 U/[micro]l) ligase (Taq DNA ligase
enzyme system), 1 [micro]l PCR product (100 ng/[micro]l), and 6.
Note exquisite sensitivity in cell strains derived from DNA damage sensing- or repair-deficient cells derived from patients with abnormal ATM, NBS1 and DNA ligase
In the blunt end, a DNA ligase
will join together the 3' end and the 5' end of one molecule with the 5' end and the 3' end, respectively, of another molecule.
The insert and vector were then ligated using T4 DNA ligase
following the manufacturer's protocol.
Ligation was carried out using T4 DNA Ligase
(Promega) in a 4[degrees]C overnight reaction.
The ligation mixture consists of 1 [micro]l of pUC19 vector, 9 [micro]l of 2x ligation buffer, 1 [micro]l of T4 DNA ligase
1 mg/mL BSA and 2 U T4 DNA ligase
(MBI Fermentas) were added.
Each ligation reaction (20 [micro]L) contained the restriction digestion product (10 [micro]L), 1 x T4 DNA ligase
buffer with EDTA, 0.