Phillips D J, Benson J M, Pruckler J M, Hooper W C
Hematologic Diseases Branch, Centers for Disease Control, Atlanta, Georgia 30333.
PCR Methods Appl. 1992 Aug;2(1):45-50. doi: 10.1101/gr.2.1.45.
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.
通过在最初的循环中使PCR反应有利于对靶序列的特异性,并在后续循环中持续高效扩增该序列,我们能够提高扩增的ras和p53序列的特异性和数量。当肿瘤细胞系和组织的DNA中存在点突变时,特异性产物纯度和产量的提高有利于增强使用直接测序和单链构象多态性(SSCP)分析进行的PCR后结果评估和解读。