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A simple method to overcome the inhibitory effect of heparin on DNA amplification

Cellular Oncology : Volume 38, Issue 5, October 2015

Cellular Oncology : Volume 38, Issue 4, August 2015

Cellular Oncology : Volume 38, Issue 3, June 2015

Cellular Oncology : Volume 38, Issue 2, April 2015

Cellular Oncology : Volume 38, Issue 1, February 2015

Cellular Oncology : Volume 37

Cellular Oncology : Volume 36

Cellular Oncology : Volume 35

Cellular Oncology : Volume 34

A simple method to overcome the inhibitory effect of heparin on DNA amplification

Content Provider	Springer Nature Link
Author	Vaughan Shaw, P. G. Walker, M. Ooi, L. Gilbert, N. Farrington, S. M. Dunlop, M. G.
Copyright Year	2015
Abstract	Genetic material from large patient cohorts is increasingly central to translational genetic research. However, patient blood samples are a finite resource and their supply and storage are often dictated by clinical and not research protocols. Our experience supports difficulty in amplifying DNA from blood stored in herparin; a scenario that other researchers may have or will encounter. This technical note describes a number of simple steps that enable successful PCR amplification.DNA was extracted using the Illustra Nucleon Genomic DNA Extraction Kit. PCR amplification was attempted using a number of commercially available PCR mastermixes.PCR DNA amplification failed using ReddyMix™ PCR Master Mix, Thermo-Start® (Thermo Scientific Inc. US) and ZymoTaq™ (Zymo research, US) PCR mastermixes, as demonstrated absence of products on gel electrophoresis. However, using the Invitrogen™ (Thermo Scientific Inc., US) Platinum® Taq DNA Polymerase, PCR products were identified on a 1% agarose gel for all samples. PCR products were cleaned with ExoSAP-IT® (Affymetrix Inc., US) and a sequencing reaction undertaken using a standard Big Dye protocol. Subsequent genotyping was successful for all samples for alleles at the CDH1 locus.From our experience a standard phenol/chloroform purification and using the Invitrogen™ Platinum® Taq has enabled the amplification of whole blood samples taken into lithium heparin and stored frozen for up to a month. This simple method may enable investigators to utilise blood taken in lithium heparin for DNA extraction and amplification.
Starting Page	493
Ending Page	495
Page Count	3
File Format	PDF
ISSN	22113428
Journal	Cellular Oncology
Volume Number	38
Issue Number	6
e-ISSN	22113436
Language	English
Publisher	Springer Netherlands
Publisher Date	2015-10-29
Publisher Institution	International Society for Cellular Oncology
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Single nucleotide polymorphism Polymerase chain reaction Heparin Cancer Research Biomedicine general Pathology Oncology
Content Type	Text
Resource Type	Article
Subject	Cancer Research Molecular Medicine Oncology

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