Open AccessPCR regimen for enhanced specificity and yield of targeted genomic DNA sequences: ras and p53.
Author(s) -
D. J. Phillips,
J M Benson,
Janet Pruckler,
W. Craig Hooper
Publication year - 1992
Publication title -
genome research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.556
H-Index - 297
eISSN - 1549-5469
pISSN - 1088-9051
DOI - 10.1101/gr.2.1.45
Subject(s) - biology , single strand conformation polymorphism , dna sequencing , dna , microbiology and biotechnology , multiple displacement amplification , genetics , genomic dna , digital polymerase chain reaction , point mutation , sequence (biology) , computational biology , polymerase chain reaction , mutation , gene , dna extraction
By weighting the PCR reaction in favor of specificity for the target sequence in the beginning cycles and for continued efficient amplification of the sequence into later cycles, we were able to show an improvement in the specificity and quantity of amplified ras and p53 sequences. Increased purity and yield of specific products favorably enhanced post-PCR evaluation and interpretation of results using direct sequencing and single-stranded conformation polymorphism (SSCP) analysis when point mutations were present in DNA from tumor cell lines and tissues.
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