I am trying for RNA isolation from fresh and frozen blood with TRIzol and QIAGEN kit but not getting very good RNA. Need suggestions!!!!
I did it with fresh blood the qiagen kit but got very few amount of RNA. However the quality was for me ok
It is better to isolate RNA from fresh samples to avoid effect of RNAase, but if nessary to isolate from fozen sample: magnisum salt used to inhibit RNAase, also RNAase free water,
Dear Sumeet Jain,
I assume you are speaking of EDTA blood in which you than use whole blood RNA isolation techniques of Trizol and/or Qiagen. In both cases, already collecting blood via such methods might cause a huge variation in your gene expression profiles after isolation, as you already will enter variance of time from withdrawal to isolation. Than using frozen blood will certainly will cause your RNA quality to be very bad in which RIN values will be much too low. Therefore, if you still have the possibility, you should use methods like PAXgene Blood RNA tubes (http://www.preanalytix.com/products/blood/RNA/paxgene-blood-rna-tube) that will preserve your RNA intact, even if you do not isolate RNA immediately and freeze the blood samples.
There are also other companies for such tubes as:
ThermoFisher https://www.thermofisher.com/order/catalog/product/4342792
NORGEN https://norgenbiotek.com/product/blood-rna-preservative-tubes
Biometrica http://biomatrica.com/rnagardblood_tube.php
There are also some publications comparing some of the above. E.g. http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0113298
Nevertheless, there are also some publications comparing different methods of isolating RNA from frozen EDTA-blood tubes. E.g. http://www.springerplus.com/content/3/1/76
Indeed, there may be a problem within the blood sampling. The processing of blood collected in common tubes (e.g. with EDTA) should be done as rapidly as possible, or you can consider a collection into stabilization tubes (e.g. Tempus Blood RNA tube) or Streck tubes for isolation of plasma. Haemolysis may also negatively affect the quality of samples. Make sure that you are working in RNAse-free environment as well. Good luck.
i think you mean the whole blood collected in EDTA ,,if it is the case i have used Qiagen and Metabion kits and they worked fine for me with good quality
Good luck
Dear Sumeet,
Have a look at this:
https://www.ncbi.nlm.nih.gov/pubmed/24567882
Good luck,
Hamid
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