Open AccessHigh-throughput SNP genotyping by combining exonuclease III, nuclease S1, and acridine-bearing PNA
Author(s) -
Bing Ren
Publication year - 2004
Publication title -
nucleic acids symposium series
Language(s) - English
Resource type - Journals
eISSN - 1746-8272
pISSN - 0261-3166
DOI - 10.1093/nass/48.1.183
Subject(s) - nuclease , dna , exonuclease , peptide nucleic acid , genotyping , chemistry , snp genotyping , microbiology and biotechnology , nucleic acid , molecular inversion probe , acridine , exonuclease iii , biochemistry , biology , genotype , dna polymerase , gene , escherichia coli , organic chemistry
Recently we reported a precise and straightforward method for genotyping of single nucleotide polymorphisms (SNPs) in double-stranded DNA substrates. The substrates were consecutively treated by two enzymes (exonuclease III and nuclease S1) in the presence of peptide nucleic acid (PNA) probes, and the resultant DNA fragments were analyzed by mass spectroscopy. Here, the selectivity of DNA scission by nuclease S1 has been greatly improved by attaching an acridine to the PNA probes. Modification at the C-terminus was especially effective. The number of DNA fragments formed has been greatly decreased so that double-stranded DNA substrates can be genotyped by using the two enzymes still more easily and precisely.
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