Affordable Access

Amplification of DNA from whole blood.

Authors
  • Burckhardt, J
Type
Published Article
Journal
PCR methods and applications
Publication Date
Feb 01, 1994
Volume
3
Issue
4
Pages
239–243
Identifiers
PMID: 8173513
Source
Medline
License
Unknown

Abstract

A method is described for the amplification by PCR of human chromosomal DNA sequences from whole blood samples. Various amounts of blood samples, with either EDTA, citrate, or heparin used as the anticoagulant, have been used to determine optimal PCR conditions for each type of sample. Up to 80% (vol/vol) of whole blood sample is tolerated in PCR with Taq polymerase. Amplification from whole blood requires the optimization of salt (K+ and Mg++) according to sample volume and type of anticoagulant used. Pretreatment of fresh blood samples to lyse the leukocytes is required for EDTA-treated blood samples and is beneficent in PCR with heparin- and citrate-treated blood samples to obtain maximal amplicon amounts. A satisfactory method of pretreating samples is freeze/thawing. In addition, EDTA-treated blood samples require a heat treatment before PCR for maximal amplicon synthesis. It appears that purification of the DNA is not necessary for any of the whole blood samples analyzed by PCR. Results of amplification reactions from unpurified hepatitis B virus (HBV) sequences of whole sera samples are shown also.

Report this publication

Statistics

Seen <100 times