Bindon C, Martindale J, Mitchell C
Department of Paediatric Oncology, Oxford Radcliffe Hospital, Oxford, UK.
Nucleic Acids Res. 1998 Jul 1;26(13):3305-8. doi: 10.1093/nar/26.13.3305.
We describe a method for producing specific PCR primers directly from PCR product, bypassing the usual need to know the primer sequence. Lack of abundance of primers derived from a PCR product is compensated for by the incorporation of an arbitrary 5'TAG sequence which acts as a surrogate template target for the bulk amplification phase. We use the technique to amplify clonospecific rearranged immunoglobulin genes, which have applications as markers of lymphoid neoplasms for tracing the success of therapy. The principle may have wider application wherever conserved and variable regions of DNA are juxtaposed.
我们描述了一种直接从PCR产物中生成特异性PCR引物的方法,无需像往常一样知道引物序列。通过掺入一个任意的5'TAG序列来补偿从PCR产物衍生的引物数量不足的问题,该序列在大量扩增阶段充当替代模板靶点。我们使用该技术扩增克隆特异性重排的免疫球蛋白基因,这些基因可作为淋巴肿瘤的标志物用于追踪治疗效果。只要DNA的保守区和可变区相邻,该原理可能具有更广泛的应用。