Linearization and purification of BAC DNA for the development of transgenic mice

被引：23

作者：

Chrast, R

Scott, HS

Antonarakis, SE

机构：

[1] Univ Geneva, Sch Med, Dept Med Genet, Lab Human Mol Genet, CH-1211 Geneva 4, Switzerland

[2] Univ Geneva, Sch Med, Grad Program Cellular & Mol Biol, CH-1211 Geneva, Switzerland

[3] Cantonal Hosp Geneva, Div Med Genet, Geneva, Switzerland

来源：

TRANSGENIC RESEARCH | 1999年 / 8卷 / 02期

关键词：

transgenic mice; BAC; protocol; linearization; purification; terminase; RARE;

D O I：

10.1023/A:1008858014473

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Bacterial artificial chromosome (BAC) vectors are increasingly used for generation of transgenic mice due to the relatively large size and the stability of their inserts compared to YACs. We have compared methods for purification and linearization of BACs, and describe an optimised protocol for preparation of high quality linear BAC DNA based on lambda terminase digestion, electroelution of linearized DNA together with simple preliminary multiplex PCR screening to detect transgenic mice. Linearized BAC DNA purified this way was successfully used for the development of transgenic mice containing 2-4 copies of the transgene.

引用

收藏

页码：147 / 150

页数：4

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