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PubMed · 1477670

Improved blood sample processing for PCR.

D Casareale·R Pottathil·R Diaco
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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.

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Acquired Immunodeficiency SyndromeBlood PreservationBuffersDNA, ViralEndopeptidase KHIVHIV SeropositivityHTLV-I InfectionsHTLV-II InfectionsHuman T-lymphotropic virus 1Human T-lymphotropic virus 2HumansLymphocytesPolymerase Chain ReactionSensitivity and SpecificitySerine EndopeptidasesSolventsSpecimen HandlingComparative StudyJournal Article

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D Casareale, R Pottathil, R Diaco. 1992. Improved blood sample processing for PCR.. https://doi.org/10.1101/gr.2.2.149

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