必须要开这个系列了,而且要对实操有一定的理解,没一篇我都会咨询实验成功过的人士,以确保未来我能带领小弟们做出来。
未来要开的系列:
Cancer模型系列:
- Cell line (DepMap)
- Mouse (JAX)
- Organoid
- Patient sample(非常难)
基础实验系列:
- WB
- RT-PCR
- co-IP
基因表达调控系列:
- 基因表达调控 - RNAi(shRNA siRNA)
- 基因表达调控 - CRISPR/Cas9
- 基因表达调控 - 过表达 CDS
- 基因表达调控+标记 - Cre/LoxP
NGS系列:
- RNA-seq
- ChIP-seq
- ATAC-seq
- Cut&Run
- scRNA-seq
- scRNA-seq/scATAC-seq
- Perturb-seq
- Spatial transcriptome
目的:
- 只要是生信和实验,通过in silico或者screening找到了hit,那肯定是要验证的,过表达实验很难,那就down吧,RNAi是首选,所以要把siRNA或者shRNA放进质粒载体,去小鼠或者其Organoid里测试。
Molecular cloning refers to the isolation of a DNA sequence from any species (often a gene), and its insertion into a vector for propagation, without alteration of the original DNA sequence.
质粒选择:
参考:
- Molecular cloning overview - techniques & workflow - YT
- pLKO-1 Vector and shRNA Design Frequently Asked Questions
- Mammalian shRNA Tools for RNAi
标签:shRNA,系列,Cancer,seq,cloning,表达,Molecular From: https://www.cnblogs.com/leezx/p/17150023.html