[Preparing of a ssDNA target in a single-round PCR with low-melt excess primer for hybridization with microarrays]

Mol Biol (Mosk). 2011 Mar-Apr;45(2):267-71.
[Article in Russian]

Abstract

Method of ssDNA preparing in single-round PCR for microarray application is described. The approach is exemplified on genotyping of DARC gene. It is opposed to two-round PCR that consists of separate symmethric and asymmethric stages. Implementation of reaction in single round is achieved by means of low-melt excess internal primer application. The primer do not anneal during symmethric stage but after decreasing of annealing temperature on asymmethric stage. The results indicate effective oligonucleotide microarray genotyping. The approach reduces time requirements and risk of contamination.

MeSH terms

  • Alleles
  • DNA Primers / chemistry
  • DNA, Single-Stranded / biosynthesis
  • DNA, Single-Stranded / genetics
  • DNA, Single-Stranded / isolation & purification*
  • Duffy Blood-Group System / blood
  • Humans
  • Nucleic Acid Hybridization
  • Oligonucleotide Array Sequence Analysis*
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Single Nucleotide / genetics
  • Receptors, Cell Surface / blood

Substances

  • ACKR1 protein, human
  • DNA Primers
  • DNA, Single-Stranded
  • Duffy Blood-Group System
  • Receptors, Cell Surface