Improved blood sample processing for PCR.
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.
Casareale
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Roche Molecular Systems, Branchburg, New Jersey 08876-1760.
Pottathil
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Diaco
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D000163:Acquired Immunodeficiency Syndrome / Q000097:blood / Q000382:microbiology; D001793:Blood Preservation; D002021:Buffers; D004279:DNA, Viral / Q000097:blood* / Q000302:isolation & purification; D019286:Endopeptidase K; D006678:HIV / Q000302:isolation & purification; D006679:HIV Seropositivity / Q000097:blood / Q000382:microbiology; D015490:HTLV-I Infections / Q000097:blood / Q000382:microbiology; D015491:HTLV-II Infections / Q000097:blood / Q000382:microbiology; D015368:Human T-lymphotropic virus 1 / Q000302:isolation & purification; D015367:Human T-lymphotropic virus 2 / Q000302:isolation & purification; D006801:Humans; D008214:Lymphocytes / Q000737:chemistry* / Q000382:microbiology; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012697:Serine Endopeptidases; D012997:Solvents; D013048:Specimen Handling
D002021:Buffers; D004279:DNA, Viral; D012997:Solvents; D012697:Serine Endopeptidases; D019286:Endopeptidase K