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DNA ligases catalyze the strand joining reaction at a nick junction.
We are currently studying the joining reaction with purified proteins to determine how the DNA double-strand breaks are repaired.
Cre utilizes the Flp, rather than topoisomerase, strategy to protect the integrity of the strand joining reaction.
In most reactions, the 5'-OH on the bottom strand of the half-site was blocked by phosphorylation (Figure 2B) to prevent the strand joining reaction.
Protection of the joining reaction from water during Cre and Flp recombination appears to be conferred by the compact design of their synaptic structures (see Figure 1A, B).
We have shown that within the resolution limits of our measurement the excision in Arabidopsis of a Ds element from a homologous chromosome does not provoke interaction of the broken chromosome with its intact homologue, and that the repair mechanism resembles the immunoglobulin V(D J joining reaction of vertebrates.
PEG 8000 appeared to increase the rate of the strand joining reaction for all variants tested.
In addition, in the extract-based end joining reaction, pol λ is able to extend an 8-oxodG-terminated primer.
In contrast to other protocols, a longer recovery time of 2 h was necessary to complete the end joining reaction before antibiotic selection was applied.
Another problem is this technique's complicated joining reaction in which a promoter cassette, the purified V gene fragment and a terminator cassette must be assembled in a specific order based on their short homology overlaps.
A-NHEJ is a evolutionarily conserved repair pathway in which double strand break repair products display short tracks of microhomology at the repair junctions and the joining reaction utilizes DNA Ligase III [ 24].
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