Question ??? RNA-Isolation fromblood - (Aug/21/2001 )
I would like to isolate RNA from blood.Can anyone give me an advice in handling with RNA and blood.What should I do, if I get the blood from the patient. Can I store it ?
You can make use of a particular Qiagen kit for purification of RNA from blood.
I am using a kit called EZNA Blood RNA kit from Omega Bio-tek, Inc. and works very well for RNA extraction from blood samples taken from patients.Here is a synopsis of the kit which I had downloaded from their official website - I hope this helps:
E.Z.N.A.® Blood RNA Kits are designed for isolation of total intracellular RNA from up to 1 ml of fresh, whole blood treated with any common anticoagulant such as heparin, EDTA, or acid-citrate-dextrose. One ml of blood typically yields 1–5 µg of total RNA. The procedure completely removes contaminants and enzyme inhibitors making total RNA isolation fast, convenient, and reliable. There is no need for phenol/chloroform extractions, and time-consuming steps such as CsCl gradient ultracentrifugation, and precipitation with isopropanol or LiCl, are eliminated. The kit is also suitable for isolation of total RNA from cultures cells, tissues, and from RNA viruses from serum or plasma. The Kits use the reversible binding properties of a new silica-based material. This is combined with the speed of mini-column spin technology. Red blood cells are selectively lysed and white cells collected by centrifugation. After lysis of white blood cells under denaturing conditions that inactivate RNases, total RNA is purified on the spin column. A specifically formulated high salt buffer system allows RNA molecules greater than 200 bases to bind to the matrix. Cellular debris and other contaminants (such as hemoglobin) are effectively washed away and high quality RNA is finally eluted in DEPC-treated sterile water.
THe kits involve spin-column technology, you do not have to mess with organic solvents and saves me alot of time. I also did some comparison shopping and while many use Qiagen kits(generally genomic RNA extractions)this is more specialized for blood and at half the cost, I got great results!!!You can check out their website (I think its firstname.lastname@example.org)for more info. Good Luck!!!