Dear collegues,
I have saved blood in EDTA tubes and saved at -80 degree. What is the experience with isolation of RNA from this frozen EDTA blood for microarray??.
will be thankful for the suggestions/comments
thank you ali for taking time....Do you have any suggestion, how we can analyze gene expression on theses EDTA samples?.
Hi, Dr. S. Muhammad
First of all, if you want to extract RNA from blood, collect the sample in RNA preservation fluid containing vials and then store it in -80. If RNA preservation fluid is unavailable, you can store the sample at 4 C for atleast 2 days; also in such case, never store the sample in -80 or -20 C as it can cause cell lysis and compromise your RNA integrity as well as yield.
You have to first isolate the WBCs from the blood sample. For that, treat your sample with RNA lysis buffer (RLB). You can find the composition online. Then centrifuge the samples to pellet down the WBCs. Also wash the cells using PBS.
you can follow the link http://www.sciencepub.net/nature/0603/08_mahongbao_TRIzol.pdf for the rest of the protocol.
Usually 7-8 ml of blood gives a RNA yield of around 30-35 microgram which is more then sufficient for various purposes. Also the quality is very good with 280/260 ratio b/w 1.95-2.1.
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