Huang S H
Department of Pediatrics, University of Southern California, Children's Hospital Los Angeles 90027.
Mol Biotechnol. 1994 Aug;2(1):15-22. doi: 10.1007/BF02789286.
The conventional polymerase chain reaction (PCR) requires that DNA sequences at both ends of the region to be amplified be known. Inverse PCR (IPCR) and anchored PCR overcome this limitation and amplify flanking unknown DNA sequences by utilizing inverse amplification and a universal primer, respectively. The major advantage of IPCR is that two gene-specific primers are reserved for specific and efficient amplification of the unknown cDNA ends on the basis of a small stretch of known sequence. The protocol consists of five steps: reverse transcription, synthesis of second strand cDNA, circularization of double strand cDNA, reopen the circle DNA, and amplification of the inverse DNA fragment.
传统的聚合酶链反应（PCR）要求待扩增区域两端的DNA序列已知。反向PCR（IPCR）和锚定PCR分别通过利用反向扩增和通用引物克服了这一限制，并扩增侧翼未知DNA序列。IPCR的主要优点是基于一小段已知序列保留两个基因特异性引物，用于特异性和高效地扩增未知cDNA末端。该方案包括五个步骤：逆转录、第二链cDNA合成、双链cDNA环化、重新打开环状DNA以及反向DNA片段的扩增。
