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Real-time PCR-based genotyping from whole blood using Taq DNA polymerase and a buffer supplemented with 1,2-propanediol and trehalose.

Authors :
Utekal P
Kocanda L
Matousek P
Wagner P
Bugajev V
Draber P
Source :
Journal of immunological methods [J Immunol Methods] 2015 Jan; Vol. 416, pp. 178-82. Date of Electronic Publication: 2014 Oct 08.
Publication Year :
2015

Abstract

Amplification of DNA templates from whole blood with Taq DNA polymerase remains a difficult task worldwide. Using a real-time PCR setup and a buffer supplemented with 1M 1,2-propanediol, 0.2M trehalose, and SYBR green I we show a reliable technique for genotyping in mice and detection of single-nucleotide polymorphisms/mutations in humans. Elimination of DNA extraction and use of the common Taq DNA polymerase and DNA dye bring about substantial savings in labor and cost.<br /> (Copyright © 2014 Elsevier B.V. All rights reserved.)

Details

Language :
English
ISSN :
1872-7905
Volume :
416
Database :
MEDLINE
Journal :
Journal of immunological methods
Publication Type :
Academic Journal
Accession number :
25305365
Full Text :
https://doi.org/10.1016/j.jim.2014.09.006