产品用途

噬菌体表面展示系统辅助噬菌体

产品特性

产生用于测序和突变的单链噬菌粒DNA

产品简介

VCSM13是M13KO7的衍生物，最初由Stratagene商业化，常用于噬菌体展示。VCSM13的表型尚未发表，确切序列尚不清楚。为了繁殖和扩增VCSM13辅助噬菌体，需要使用TG1、ER2738或菌株XL1 Blue MRF′大肠杆菌菌株，因为这些菌株含有supE44突变，该突变提供谷氨酰胺抑制剂tRNA。VCSM13辅助噬菌体严格用于从已切除的噬菌体中产生单链DNA（这也是M13KO7辅助噬菌体的用途）。

规格

500ul/管，1管

贮存

Recommends storing them at -80℃ with 50% Glycerol. As long as there is a -80℃ stock in the lab,   the amplified lab prep can be stored at +4℃.

扩增

1. Streak out TG1 onto an 2YT plate.

2. Pick a single colony of TG1 and inoculate into 5ml 2×YT and grow until OD600 = 0.3 (~2.5×10⁸ cells/ml).

3. Add helper phage at a multiplicity of infection (MOI) of 20:1 (phage-to-cells ratio).

Note: If amplifying VCSM13 helper phage, add Kanamycin to a final concentration of 25μg/ml to the medium 30 minutes after the helper phage and cells have been allowed to grow together.

4. Grow the culture at 37℃ with vigorous aeration (~300rpm) for 8 hours.

5. Heat the culture to 65℃ for 15 minutes.

6. Spin down the cell debris and transfer the supernatant to a fresh tube.

7. Titer the helper phage produced.