Bates G P, Wainwright B J, Williamson R, Brown S D
Mol Cell Biol. 1986 Nov;6(11):3826-30. doi: 10.1128/mcb.6.11.3826-3830.1986.
A bank of cloned DNA sequences from the distal half of the short arm of human chromosome 2 was generated by using microdissection and microcloning techniques. DNA was purified from 106 chromosomal fragments, manually dissected from peripheral lymphocytes in metaphase, and cloned into the EcoRI site of lambda gt10. A total of 257 putative recombinants were recovered, of which 41% were found to contain human inserts. The mean insert size was 380 base pairs (median size, 83 base pairs), and fewer than 10% of the clones contained highly repetitive sequences. All single-copy sequences examined were shown to map to the short arm of chromosome 2 by using hybrid panels. This technique provides a rapid method of isolating probes specific to a human subchromosomal region to generate linked markers to genetic diseases for which the chromosomal location is known.
通过显微切割和微克隆技术构建了一组来自人类2号染色体短臂远端一半的克隆DNA序列。从106个染色体片段中纯化DNA，这些片段是从处于中期的外周淋巴细胞中手工切割得到的，然后克隆到λgt10的EcoRI位点。共回收了257个推定重组体，其中41%被发现含有人类插入片段。平均插入片段大小为380个碱基对（中位数大小为83个碱基对），少于10%的克隆含有高度重复序列。通过使用杂交板，所有检测的单拷贝序列都被证明定位于2号染色体的短臂。该技术提供了一种快速方法，用于分离特定于人类亚染色体区域的探针，以生成与已知染色体位置的遗传疾病相关的连锁标记。
