Author:
Casareale D,Pottathil R,Diaco R
Abstract
To simplify our procedure for blood sample processing for PCR, we introduced a simpler, shorter, and more cost-effective method for the separation of peripheral blood lymphocytes (PBL) and DNA extraction for amplification with Taq polymerase. By this method, blood samples are processed in two simple 15-min steps: (1) separation of PBLs from whole blood by red blood cell lysis with the Roche Specimen Washing Solution, and (2) DNA extraction by heat-detergent treatment of separated PBLs. This new method is simpler than the standard Ficoll-Hypaque method for PBLs separation and Proteinase K digestion for DNA extraction. It is not inhibitory to DNA amplification and it allows effective processing of blood samples even after prolonged storage (as long as 8 days) at room temperature.
Publisher
Cold Spring Harbor Laboratory
Subject
Genetics (clinical),Genetics
Reference11 articles.
1. Mullis, K.B., Faloona, F.A. Scharf, S.J. Saiki, R.K. Horn, G.T. and Erlich. H.A. 1986. Specific enzymatic amplification of DNA in vitro: The polymerase chain reaction. Cold Spring Harbor Symp. Quant. Biol. 51:
2. Isolation of mononuclear cells and granulocytes from human blood.;Scand. J. Clin. Lab. Invest.,1968
3. Micromethod for rapid separation of lymphocytes from peripheral blood.;Ann. Clin. Lab. Sci.,1971
4. Ausubel, F.M. 1987. Current protocols in molecular biology. Wiley, New York.
5. A general method for isolation of high molecular weight DNA from eukaryotes
Cited by
5 articles.
订阅此论文施引文献
订阅此论文施引文献，注册后可以免费订阅5篇论文的施引文献，订阅后可以查看论文全部施引文献