Abstract:
:We present a method that allows preparing long DNA containing defined mismatches without the use of gel electrophoretic or chromatographic purification steps. The preparation starts with the synthesis of two PCR products, which are identical except for those positions that will later form the mismatches. One of the PCR primers must be 5'-phosphorylated, such that in two separate reactions two PCR products are obtained, which are 5'-phosphorylated in one strand. After removal of the phosphorylated strands by lambda-exonuclease, the resulting single strands are hybridized to form the mismatch-containing heteroduplex. The application of this procedure is demonstrated for the analysis of the Escherichia coli MutHLS system.
journal_name
Biol Chemjournal_title
Biological chemistryauthors
Thomas E,Pingoud A,Friedhoff Pdoi
10.1515/BC.2002.166keywords:
subject
Has Abstractpub_date
2002-09-01 00:00:00pages
1459-62issue
9eissn
1431-6730issn
1437-4315journal_volume
383pub_type
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