Hadano S, Ikeda J E
Ikeda GenoSPHERE Project, ERATO, JRDC.
Nihon Rinsho. 1993 Sep;51(9):2240-5.
We have developed an argon ion laser chromosome microdissection technique in conjunction with a single unique primer polymerase chain reaction (SUP-PCR) to directly amplify microdissected chromosomes. The 22-mer primer used in PCR, although unique in sequence, randomly primed and amplified any target DNA. These methods were applied to both the terminal region of the human chromosome 4p (4p 16) and Xq (Xq26-q28), and two chromosome region-specific DNA libraries were constructed. The resulting libraries contained approximately 1000 nonoverlapping DNA sequences with an average size of 230-350 bp, at an average spacing of 10-65 Kbp along the chromosomes of origin. Our new method is a simple and general approach for constructing a chromosome region-specific DNA library from a single metaphase spread.
我们开发了一种氩离子激光染色体显微切割技术，并结合单独特异性引物聚合酶链反应（SUP-PCR）来直接扩增显微切割的染色体。PCR中使用的22聚体引物，尽管序列独特，但能随机引发并扩增任何目标DNA。这些方法应用于人类染色体4p（4p16）和Xq（Xq26-q28）的末端区域，并构建了两个染色体区域特异性DNA文库。所得文库包含约1000个非重叠DNA序列，平均大小为230-350bp，沿原始染色体的平均间距为10-65Kbp。我们的新方法是一种从单个中期铺片中构建染色体区域特异性DNA文库的简单通用方法。
