Polymerase chain reaction amplification of specific alleles: a general method of detection of mutations, polymorphisms, and haplotypes.

摘要: Publisher Summary The polymerase chain reaction (PCR) utilizes two oligonucleotide primers to amplify a segment of DNA more than 1 million-fold. PCR can be adapted for the rapid detection known single-base changes in by using specially designed oligonucleotides amplification specific alleles (PASA). principle this method is design an primer that will preferentially one allele over another. PASA generally applicable technique point mutations or polymorphisms and presence small deletions insertion. But has certain disadvantages are eliminated another technique, multiple (PAMSA). This allows single reaction. PAMSA useful detecting polymorphisms. chapter discusses general haplotypes. been used perform population screening, haplotype analysis, patient carrier testing. Haplotypes genetics medicine. However, determining linkage haplotypes absence samples from appropriate family members difficult laborious. provide reproducible haplotyping individual relatives. method, termed “double PASA,” uses four pairs allele-specific differentially each possible biallelic Double important tool doubly heterozygous individuals because physical on strand necessary determine haplotype.