Shimada H, Tada Y
Mitsui Plant Biotechnology Research Institute, Tsukuba, Japan.
Gene. 1991 Feb 15;98(2):243-8. doi: 10.1016/0378-1119(91)90180-j.
Polymerase chain reaction (PCR) is an efficient method for obtaining a desired nt sequence if both required primers can hybridize to the target DNA molecule specifically. A rapid and simple PCR-based method for analyzing plasmids using intact cells was established. An attempt to target a rice waxy sequence by PCR using homologous primers was also carried out. In three cases, specific fragments were amplified and their nucleotide sequences were determined. However, the cloned rice waxy gene contained base substitution mutations. The cumulative frequency of mutation after 30 polymerization cycles was estimated to be one in 500 bp.
如果所需的两种引物都能与目标DNA分子特异性杂交，聚合酶链反应（PCR）就是获得所需核苷酸序列的有效方法。建立了一种基于PCR的快速简便的方法，用于使用完整细胞分析质粒。还尝试使用同源引物通过PCR靶向水稻蜡质序列。在三种情况下，扩增出了特异性片段并测定了它们的核苷酸序列。然而，克隆的水稻蜡质基因包含碱基替换突变。估计在30个聚合循环后的突变累积频率为每500个碱基对中有一个。
