LOW-FREQUENCY CHIMERIC YEAST ARTIFICIAL CHROMOSOME LIBRARIES FROM FLOW-SORTED HUMAN CHROMOSOME-16 AND CHROMOSOME-21

被引：43

作者：

MCCORMICK, MK ^{[1
]}

CAMPBELL, E ^{[1
]}

DEAVEN, L ^{[1
]}

MOYZIS, R ^{[1
]}

机构：

[1] LOS ALAMOS NATL LAB,CTR HUMAN GENOME STUDIES,LOS ALAMOS,NM 87544

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1993年 / 90卷 / 03期

关键词：

D O I：

10.1073/pnas.90.3.1063

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Construction of chromosome-specific yeast artificial chromosome (YAC) libraries from sorted chromosomes was undertaken (i) to eliminate drawbacks associated with first-generation total genomic YAC libraries, such as the high frequency of chimeric YACs, and (ii) to provide an alternative method for generating chromosome-specific YAC libraries in addition to isolating such collections from a total genomic library. Chromosome-specific YAC libraries highly enriched for human chromosomes 16 and 21 were constructed. By maximizing the percentage of fragments with two ligatable ends and performing yeast transformations with less than saturating amounts of DNA in the presence of carrier DNA, YAC libraries with a low percentage of chimeric clones were obtained. The smaller number of YAC clones in these chromosome-specific libraries reduces the effort involved in PCR-based screening and allows hybridization methods to be a manageable screening approach.

引用

页码：1063 / 1067

页数：5

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