We are limited in our material, primary stroma cells from patients (300.000-500.00cells/T25) and we need RNA from this cells for a deep sequencing study. Someone can give us some advice which Qiagen (or other company) kit will give good results? Until now the cells are in culture or frozen (we have to defrost and culture them).
Thanks a lot
I would highly recommend the kit Cat No./ID: 74104
RNeasy Mini Kit because of the high purity and performance yield of RNA amount per sample. Even tough, considering your cells amount, the traditional TRIZOL protocol would also work with a lower purity level. http://ycga.yale.edu/microarrays/protocols/TRIZOLRNAIsolation_092107_tcm240-21453_tcm240-284-32.pdf
Good luck!
Thanks for helping us! I will have a look at the side for more details.
Do you know if it´s including also a step to remove genomic DNA?
RNeasy kits recommend 'on-column' DNase treatment (also provided by Qiagen),...I think this may be provided with the RNeasy kit. Alternatively you can DNase treat extracts using any number of commercially available RNase free DNA kits.
I've used both approaches and both do the job. I personally prefer post extraction DNase treatment
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