丁香实验_LOGO
登录
提问
我要登录
|免费注册
点赞
收藏
wx-share
分享

制备BAC用DNA

互联网

2069

Preparation of BAC (Bacterial Artificial Chromosome) DNA with CONCERT ™ High Purity Plasmid Purification System1 Courtesy Lisha Xu and Alice C. Young, Molecular and Cell Biology Research and Development, Life Technolgies, Inc., Rockville, MD 20849.

Before Beginning:

Prepare a 20 h culture of BAC containing bacteria in 2X YT and appropriate antibiotic. The OD600 of the final culture should be 5.0 -0.5.

Add RNAse A to E1 to a final concentration of 400 µg/mL.

Increase NaCl concentration in Wash Buffer (E5) from 0.8 M to 0.9 M NaCl by adding 0.58 g NaCl per 100 mL E5. Conductivity should be 72 mS. This increase in salt will reduce the RNA contamination in the BAC prep.

Pre-warm Buffer E6 to 50℃.

NOTE: In this application, we recommend 400 µg RNAse A/mL of Suspension Buffer (E1) because of the extremely low copy number of BACs.

Equilibrate the column with Equilibration Buffer (Buffer E4) Allow the solution.

提问
扫一扫
丁香实验小程序二维码
实验小助手
丁香实验公众号二维码
扫码领资料
反馈
TOP
打开小程序