THE DEVELOPMENT AND APPLICATION OF AUTOMATED GRIDDING FOR EFFICIENT SCREENING OF YEAST AND BACTERIAL ORDERED LIBRARIES

被引：70

作者：

BENTLEY, DR ^{[1
]}

TODD, C ^{[1
]}

COLLINS, J ^{[1
]}

HOLLAND, J ^{[1
]}

DUNHAM, I ^{[1
]}

HASSOCK, S ^{[1
]}

BANKIER, A ^{[1
]}

GIANNELLI, F ^{[1
]}

机构：

[1] MRC, MOLEC BIOL LAB, CAMBRIDGE, ENGLAND

来源：

GENOMICS | 1992年 / 12卷 / 03期

关键词：

D O I：

10.1016/0888-7543(92)90445-X

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

An automated gridding procedure for the inoculation of yeast and bacterial clones in high-density arrays has been developed. A 96-pin inoculating tool compatible with the standard microtiter plate format and an eight-position tablet have been designed to fit the Biomek 1000 programmable robotic workstation (Beckman Instruments). The system is used to inoculate six copies of 80 × 120-mm filters representing a total of ∼20,000 individual clones in approximately 3 h. Highdensity arrays of yeast artificial chromosome (YAC) and cosmid clones have been used for rapid large-scale hybridization screens of ordered libraries. In addition, an improved PCR library screening strategy has been developed using strips cut from the high-density arrays to prepare row and column DNA pools for PCR analysis. This strategy eliminates the final hybridization step and allows identification of a single clone by PCR in 2 days. The development of automated gridding technology will have a significant impact on the establishment of fully versatile screening of ordered library resources for genomic studies. © 1992.

引用

页码：534 / 541

页数：8

共 28 条

[1] A 3.5 GENOME EQUIVALENT MULTIACCESS YAC LIBRARY - CONSTRUCTION, CHARACTERIZATION, SCREENING AND STORAGE [J].