跳转至内容
Merck
  • Optimized protocol to create deletion in adherent cell lines using CRISPR/Cas9 system.

Optimized protocol to create deletion in adherent cell lines using CRISPR/Cas9 system.

STAR protocols (2021-11-09)
Umer Farooq, Dimple Notani
摘要

The CRISPR/Cas9 system is a powerful tool for genome editing and is adaptable for a wide range of applications. Here, we have put together a step-by-step protocol for generating knockout cell lines (coding or non-coding region) using CRISPR/Cas9 tool. The protocol below has been tested on adherent cell lines such as HeLa and MCF7. However, it may easily be adapted to other adherent cell lines with minor variations. For complete details on the use and execution of this protocol, please refer to Farooq et al. (2021).

材料
货号
品牌
产品描述

Sigma-Aldrich
琼脂糖, BioReagent, for molecular biology, low EEO
Sigma-Aldrich
聚-D-赖氨酸 氢溴酸盐, mol wt 70,000-150,000, lyophilized powder, γ-irradiated, BioReagent, suitable for cell culture
Sigma-Aldrich
氨苄西林 钠盐, powder or crystals, BioReagent, suitable for cell culture
Sigma-Aldrich
海美溴铵, ≥94% (titration)
Sigma-Aldrich
LB Broth with agar (Luria low salt), Powder microbial growth medium