Effects of a novel cell stabilizing reagent on DNA amplification by PCR as compared to traditional stabilizing reagents

Kausik Das, M. Rohan Fernando, Sara Basiaga, Stephanie M. Wigginton, Tom Williams

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Stabilization of nucleated blood cells by cell stabilizing reagent (BCT reagent) present in the Cell-Free DNA BCT™ blood collection device and consequent prevention of cell-free DNA contamination by cellular DNA during sample storage and shipping have previously been reported. This study was conducted to investigate the effect of this novel cell stabilizing reagent on DNA amplification by PCR as compared to traditional cell stabilizing reagents, formaldehyde and glutaraldehyde. A 787. bp long DNA fragment from human glyceraldehydes-3-phosphate dehydrogenase (GAPDH) gene was amplified by PCR and used as model system. DNA samples and blood samples were treated with BCT reagent, 0.1% formaldehyde or 0.1% glutaraldehyde at room temperature. DNA amplification was studied using conventional and real-time quantitative PCR. Results indicate that exposure of DNA to the BCT reagent for up to 14 days had no effect on DNA amplification by PCR as compared to the untreated control DNA. However, there was statistically significant decrease in DNA amplification in the DNA samples treated with formaldehyde and glutaraldehyde. We conclude that the BCT reagent used in Cell-Free DNA BCT blood collection device to prevent cell-free DNA contamination by cellular DNA had no effect on DNA amplification by PCR.

Original languageEnglish (US)
Pages (from-to)55-60
Number of pages6
JournalActa Histochemica
Volume116
Issue number1
DOIs
StatePublished - Jan 1 2014

Fingerprint

Polymerase Chain Reaction
DNA
Glutaral
DNA Contamination
Formaldehyde
Equipment and Supplies
Glyceraldehyde-3-Phosphate Dehydrogenases
Real-Time Polymerase Chain Reaction
Blood Cells
Temperature
Genes

Keywords

  • Cell-free DNA BCT reagent
  • DNA amplification
  • DNA fluorescence
  • Formaldehyde free stabilizing reagent

ASJC Scopus subject areas

  • Histology
  • Cell Biology

Cite this

Effects of a novel cell stabilizing reagent on DNA amplification by PCR as compared to traditional stabilizing reagents. / Das, Kausik; Fernando, M. Rohan; Basiaga, Sara; Wigginton, Stephanie M.; Williams, Tom.

In: Acta Histochemica, Vol. 116, No. 1, 01.01.2014, p. 55-60.

Research output: Contribution to journalArticle

Das, Kausik ; Fernando, M. Rohan ; Basiaga, Sara ; Wigginton, Stephanie M. ; Williams, Tom. / Effects of a novel cell stabilizing reagent on DNA amplification by PCR as compared to traditional stabilizing reagents. In: Acta Histochemica. 2014 ; Vol. 116, No. 1. pp. 55-60.
@article{33bb0e90cd2c4e06b4ea34bd2816cd39,
title = "Effects of a novel cell stabilizing reagent on DNA amplification by PCR as compared to traditional stabilizing reagents",
abstract = "Stabilization of nucleated blood cells by cell stabilizing reagent (BCT reagent) present in the Cell-Free DNA BCT™ blood collection device and consequent prevention of cell-free DNA contamination by cellular DNA during sample storage and shipping have previously been reported. This study was conducted to investigate the effect of this novel cell stabilizing reagent on DNA amplification by PCR as compared to traditional cell stabilizing reagents, formaldehyde and glutaraldehyde. A 787. bp long DNA fragment from human glyceraldehydes-3-phosphate dehydrogenase (GAPDH) gene was amplified by PCR and used as model system. DNA samples and blood samples were treated with BCT reagent, 0.1{\%} formaldehyde or 0.1{\%} glutaraldehyde at room temperature. DNA amplification was studied using conventional and real-time quantitative PCR. Results indicate that exposure of DNA to the BCT reagent for up to 14 days had no effect on DNA amplification by PCR as compared to the untreated control DNA. However, there was statistically significant decrease in DNA amplification in the DNA samples treated with formaldehyde and glutaraldehyde. We conclude that the BCT reagent used in Cell-Free DNA BCT blood collection device to prevent cell-free DNA contamination by cellular DNA had no effect on DNA amplification by PCR.",
keywords = "Cell-free DNA BCT reagent, DNA amplification, DNA fluorescence, Formaldehyde free stabilizing reagent",
author = "Kausik Das and Fernando, {M. Rohan} and Sara Basiaga and Wigginton, {Stephanie M.} and Tom Williams",
year = "2014",
month = "1",
day = "1",
doi = "10.1016/j.acthis.2013.05.002",
language = "English (US)",
volume = "116",
pages = "55--60",
journal = "Acta Histochemica",
issn = "0065-1281",
publisher = "Urban und Fischer Verlag Jena",
number = "1",

}

TY - JOUR

T1 - Effects of a novel cell stabilizing reagent on DNA amplification by PCR as compared to traditional stabilizing reagents

AU - Das, Kausik

AU - Fernando, M. Rohan

AU - Basiaga, Sara

AU - Wigginton, Stephanie M.

AU - Williams, Tom

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Stabilization of nucleated blood cells by cell stabilizing reagent (BCT reagent) present in the Cell-Free DNA BCT™ blood collection device and consequent prevention of cell-free DNA contamination by cellular DNA during sample storage and shipping have previously been reported. This study was conducted to investigate the effect of this novel cell stabilizing reagent on DNA amplification by PCR as compared to traditional cell stabilizing reagents, formaldehyde and glutaraldehyde. A 787. bp long DNA fragment from human glyceraldehydes-3-phosphate dehydrogenase (GAPDH) gene was amplified by PCR and used as model system. DNA samples and blood samples were treated with BCT reagent, 0.1% formaldehyde or 0.1% glutaraldehyde at room temperature. DNA amplification was studied using conventional and real-time quantitative PCR. Results indicate that exposure of DNA to the BCT reagent for up to 14 days had no effect on DNA amplification by PCR as compared to the untreated control DNA. However, there was statistically significant decrease in DNA amplification in the DNA samples treated with formaldehyde and glutaraldehyde. We conclude that the BCT reagent used in Cell-Free DNA BCT blood collection device to prevent cell-free DNA contamination by cellular DNA had no effect on DNA amplification by PCR.

AB - Stabilization of nucleated blood cells by cell stabilizing reagent (BCT reagent) present in the Cell-Free DNA BCT™ blood collection device and consequent prevention of cell-free DNA contamination by cellular DNA during sample storage and shipping have previously been reported. This study was conducted to investigate the effect of this novel cell stabilizing reagent on DNA amplification by PCR as compared to traditional cell stabilizing reagents, formaldehyde and glutaraldehyde. A 787. bp long DNA fragment from human glyceraldehydes-3-phosphate dehydrogenase (GAPDH) gene was amplified by PCR and used as model system. DNA samples and blood samples were treated with BCT reagent, 0.1% formaldehyde or 0.1% glutaraldehyde at room temperature. DNA amplification was studied using conventional and real-time quantitative PCR. Results indicate that exposure of DNA to the BCT reagent for up to 14 days had no effect on DNA amplification by PCR as compared to the untreated control DNA. However, there was statistically significant decrease in DNA amplification in the DNA samples treated with formaldehyde and glutaraldehyde. We conclude that the BCT reagent used in Cell-Free DNA BCT blood collection device to prevent cell-free DNA contamination by cellular DNA had no effect on DNA amplification by PCR.

KW - Cell-free DNA BCT reagent

KW - DNA amplification

KW - DNA fluorescence

KW - Formaldehyde free stabilizing reagent

UR - http://www.scopus.com/inward/record.url?scp=84894901873&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84894901873&partnerID=8YFLogxK

U2 - 10.1016/j.acthis.2013.05.002

DO - 10.1016/j.acthis.2013.05.002

M3 - Article

C2 - 23810682

AN - SCOPUS:84894901873

VL - 116

SP - 55

EP - 60

JO - Acta Histochemica

JF - Acta Histochemica

SN - 0065-1281

IS - 1

ER -