Hello everyone,
My current project involves identification of mutations in the tumours of cigarette smoke induced lung cancer mice models. I am interested in studying gene expression patterns using RNA sequencing on the tumours. Can anyone guide me as to what things I need to consider before performing RNA sequencing.
Thank you.
I agree with Nicole
I will just mention some critical point and possible trouble you will face during experiment
1- During crushing tissue you will loss some genomic RNA, so need to optimize this step
2-The temperature is very important during extraction , use always ice
3-Before runing PCR make sure your rna is well purified (nanodrop if possible)
I am sure you will find good result
Good Job!!!
First and foremost, keep the samples IMMEDIATELY after obtention in liquid nitrogen, or even better, in RNAlater at -20, which is much more practical, and allows for at least 20 de-freezing and re-freezing cycles. RNA degrades VERY fast, almost by just looking at the sample....
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