Polymer-stimulated ligation: enhanced blunt- or cohesive-end ligation of DNA or deoxyribooligonucleotides by T4 DNA ligase in polymer solutions.
The rates of blunt-end and cohesive-end ligation of DNA by T4 DNA ligase are increased by orders of magnitude in the presence of high concentrations of a variety of nonspecific polymers such as polyethylene glycol, Ficoll, bovine plasma albumin, or glycogen. Blunt-end ligation of small self-complementary oligodeoxyribonucleotides is also stimulated. The use of polyethylene glycol 6000 in such systems is characterized in some detail. Conditions are suggested which either greatly increase the rate of formation of and size of linear ligation products or which allow smaller but significant improvements in the amounts of circular ligation products.
D004247:DNA; D011088:DNA Ligases / Q000378:metabolism*; D003854:Deoxyribonucleotides / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D007700:Kinetics; D010957:Plasmids; D011092:Polyethylene Glycols / Q000494:pharmacology; D011118:Polynucleotide Ligases / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013604:T-Phages / Q000201:enzymology*
D003854:Deoxyribonucleotides; D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols; D004247:DNA; D011088:DNA Ligases; D011118:Polynucleotide Ligases