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[人YAC文库三维PCR筛选系统的建立及质量评估]

[The establishment of a three dimensional PCR screening system and quality evaluation of human YAC library].

作者信息

Ye Q, Chen S, Chen Z, Han S, Shi J, Yu L, Zhao S

机构信息

Shanghai Institute of Hematology, Rui Jin Hospital, Shanghai Second Medical University, Shanghai.

出版信息

Yi Chuan Xue Bao. 1996;23(4):249-54.

PMID:8950854
Abstract

In order to screen out YAC clones in the CEPH YAC library related to a definite gene, DNA fragment or locus, we need to establish a screening system. This paper includes the establishment of this system and the evaluation of its feasibility. According to the experiments, we suggest that it is better to screen the Mega YAC part of the library at first and put five culture plates as a group. Meanwhile, FISH experiments to assess the quality of CEPH YAC library and the first generation physical map of human genome, Were conducted FISH experiments on 26 YAC clones were carried out. There were 13 clones that were chimeric (about 50%), 6 clones that were mapped by mistake (about 23%). So, there were only 9 clones that were mapped correctly and were not chimeric (about 35%).

摘要

为了从CEPH酵母人工染色体(YAC)文库中筛选出与特定基因、DNA片段或基因座相关的YAC克隆,我们需要建立一个筛选系统。本文包括该系统的建立及其可行性评估。根据实验,我们建议首先筛选文库中的Mega YAC部分,并将五个培养板作为一组。同时,进行了荧光原位杂交(FISH)实验以评估CEPH YAC文库的质量和人类基因组的第一代物理图谱,对26个YAC克隆进行了FISH实验。其中有13个克隆是嵌合体(约50%),6个克隆定位错误(约23%)。因此,只有9个克隆定位正确且不是嵌合体(约35%)。

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