methods to extract RNA from blood sample? - (May/19/2013 )
can anybody tel the methods to extract good quality RNA from blood sample?
i have tried trizol method and some company kits but didnt get good results.
Do you need them for rt-pcr? Compared to what are you saying you didn't get good result? Trizol should be enough, just run the next step of your experiment and see. Maybe it's too early to say you don't have enough RNA.
yes i need it for RT-PCR to make cDNA. the trizol method extracts the RNA but yield is quit low which dont make cDNA for further PCR amplification.
any tips regarding to extraction u can tell?
just be careful in the last step, RNA pellet is usually transparent.
The standard method is to collect blood samples in Paxgene or Tempus vacutainers specifically for blood RNA extraction. But if your blood samples are collected in regular anticoagulants, here are some methods for RNA extraction from whole blood.
For fresh whole blood:
Prepare buffy coat then extract RNA from the the buffy coat using Trizol or other RNA extraction kits.
Lyse whole blood with AquaPreserve, remove proteins with ProSink, pellet DNA/RNA with isopropanol, and then remove DNA with DNase I.
For Frozen whole blood:
Thaw frozen whole blood in AquaPreserve, remove proteins with ProSink, pellet DNA/RNA with isopropanol, and then remove DNA with DNase I.