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PMID: 1477670 Published · ppublish English Comparative Study Journal Article

Improved blood sample processing for PCR.

PCR methods and applications ·Vol. 2 ·No. 2 ·1992-11-00 ·Pages 149-53

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.

MeSH Terms
Acquired Immunodeficiency Syndrome/blood,microbiology Blood Preservation Buffers DNA, Viral/blood,isolation & purification Endopeptidase K HIV/isolation & purification HIV Seropositivity/blood,microbiology HTLV-I Infections/blood,microbiology HTLV-II Infections/blood,microbiology Human T-lymphotropic virus 1/isolation & purification Human T-lymphotropic virus 2/isolation & purification Humans Lymphocytes/chemistry,microbiology Polymerase Chain Reaction Sensitivity and Specificity Serine Endopeptidases Solvents Specimen Handling
Chemicals
Buffers DNA, Viral Solvents Serine Endopeptidases Endopeptidase K
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Casareale D
Roche Molecular Systems, Branchburg, New Jersey 08876-1760.
Pottathil R
Diaco R
Article Info
Journal
PCR methods and applications
Abbr.
PCR Methods Appl
ISSN
1054-9803
Published
1992-11-00
Pages
149-53
Language
English
Region
United States
NLM ID
9201445
Subset
IM
Analysis Services
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