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一个3.5基因组当量的多用途酵母人工染色体文库：构建、表征、筛选及保存

A 3.5 genome equivalent multi access YAC library: construction, characterisation, screening and storage.

作者信息

Anand R, Riley J H, Butler R, Smith J C, Markham A F

机构信息

ICI Diagnostics, Northwich, Cheshire, UK.

出版信息

Nucleic Acids Res. 1990 Apr 25;18(8):1951-6. doi: 10.1093/nar/18.8.1951.

DOI:10.1093/nar/18.8.1951
PMID:2186372
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC330667/
Abstract

The construction of a yeast artificial chromosome (YAC) primary gridded library of 35,000 clones from human lymphoblastoid (48,XXXX) cell line DNA is described. The average YAC size is approximately 350kb representing a greater than 3.5 times coverage of the genome. The library is stored at -70 degrees C as gridded clones on nylon filters impregnated with 20% glycerol and as glycerol suspensions of individual clones in microtitre plates providing a prolonged multi-user potential. To date we have used 14 single copy probes to screen this library by colony hybridisation as well as PCR and have isolated between 1 and 5 YAC clones for every probe.

摘要

本文描述了一个酵母人工染色体（YAC）初级网格文库的构建，该文库包含来自人淋巴母细胞（48,XXXX）细胞系DNA的35000个克隆。YAC的平均大小约为350kb，代表基因组覆盖度大于3.5倍。该文库以网格克隆的形式保存在浸渍有20%甘油的尼龙滤膜上，保存在-70℃，同时也以单个克隆的甘油悬浮液形式保存在微量滴定板中，具有长期多用户使用的潜力。迄今为止，我们已使用14个单拷贝探针通过菌落杂交以及PCR筛选该文库，每个探针已分离出1至5个YAC克隆。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/512d/330667/844edf5d178f/nar00192-0022-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/512d/330667/ab15b4bb4598/nar00192-0020-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/512d/330667/e8d008d412aa/nar00192-0021-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/512d/330667/e06d54bf760c/nar00192-0022-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/512d/330667/844edf5d178f/nar00192-0022-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/512d/330667/ab15b4bb4598/nar00192-0020-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/512d/330667/e8d008d412aa/nar00192-0021-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/512d/330667/e06d54bf760c/nar00192-0022-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/512d/330667/844edf5d178f/nar00192-0022-b.jpg

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引用本文的文献

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A giant novel gene undergoing extensive alternative splicing is severed by a Cornelia de Lange-associated translocation breakpoint at 3q26.3.一个经历广泛可变剪接的巨大新基因被3q26.3处与科妮莉亚·德朗格综合征相关的易位断点切断。
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