Alternative RNA extraction - (Oct/28/2008 )
Can anyone please help. I have some human biopsies stored in Trizol and am extracting RNA by the phenol-chloroform method. Unfortunately my purities haven't been very good giving poor 26/230 ratios on the spectrometer. when i make cDNA and then PCR it the results are terrible. Obviously i need more practice but other people use different methods and kits which they say are quicker and more efficient.
What i want to know is:
Is there an alternative kit or method i can use given that all my samples are currently stored, frozen in Trizol. I have been told i should have stored my samples in RNAlater for this to be possible.
Novice needs help
Thanks any suggestions
Prash
Disruption and homogenization of tissue COMPLETELY is essential for RNA extraction. First get high yield of RNA and then repurify by Qiagen column. Using robust RT kit.