Digital PCR

Research output: Contribution to journalArticle

Abstract

The identification of predefined mutations expected to be present in a minor fraction of a cell population is important for a variety of basic research and clinical applications. Here, we describe an approach for transforming the exponential, analog nature of the PCR into a linear, digital signal suitable for this purpose. Single molecules are isolated by dilution and individually amplified by PCR; each product is then analyzed separately for the presence of mutations by using fluorescent probes. The feasibility of the approach is demonstrated through the detection of a mutant ras oncogene in the stool of patients with colorectal cancer. The process provides a reliable and quantitative measure of the proportion of variant sequences within a DNA sample.

Original languageEnglish (US)
Pages (from-to)9236-9241
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number16
DOIs
StatePublished - Aug 3 1999

Fingerprint

Polymerase Chain Reaction
Mutation
ras Genes
Fluorescent Dyes
Colorectal Neoplasms
DNA
Research
Population

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Digital PCR. / Vogelstein, Bert; Kinzler, Kenneth W.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, No. 16, 03.08.1999, p. 9236-9241.

Research output: Contribution to journalArticle

@article{7370d7e89ea748039e82367774f7a899,
title = "Digital PCR",
abstract = "The identification of predefined mutations expected to be present in a minor fraction of a cell population is important for a variety of basic research and clinical applications. Here, we describe an approach for transforming the exponential, analog nature of the PCR into a linear, digital signal suitable for this purpose. Single molecules are isolated by dilution and individually amplified by PCR; each product is then analyzed separately for the presence of mutations by using fluorescent probes. The feasibility of the approach is demonstrated through the detection of a mutant ras oncogene in the stool of patients with colorectal cancer. The process provides a reliable and quantitative measure of the proportion of variant sequences within a DNA sample.",
author = "Bert Vogelstein and Kinzler, {Kenneth W}",
year = "1999",
month = "8",
day = "3",
doi = "10.1073/pnas.96.16.9236",
language = "English (US)",
volume = "96",
pages = "9236--9241",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "16",

}

TY - JOUR

T1 - Digital PCR

AU - Vogelstein, Bert

AU - Kinzler, Kenneth W

PY - 1999/8/3

Y1 - 1999/8/3

N2 - The identification of predefined mutations expected to be present in a minor fraction of a cell population is important for a variety of basic research and clinical applications. Here, we describe an approach for transforming the exponential, analog nature of the PCR into a linear, digital signal suitable for this purpose. Single molecules are isolated by dilution and individually amplified by PCR; each product is then analyzed separately for the presence of mutations by using fluorescent probes. The feasibility of the approach is demonstrated through the detection of a mutant ras oncogene in the stool of patients with colorectal cancer. The process provides a reliable and quantitative measure of the proportion of variant sequences within a DNA sample.

AB - The identification of predefined mutations expected to be present in a minor fraction of a cell population is important for a variety of basic research and clinical applications. Here, we describe an approach for transforming the exponential, analog nature of the PCR into a linear, digital signal suitable for this purpose. Single molecules are isolated by dilution and individually amplified by PCR; each product is then analyzed separately for the presence of mutations by using fluorescent probes. The feasibility of the approach is demonstrated through the detection of a mutant ras oncogene in the stool of patients with colorectal cancer. The process provides a reliable and quantitative measure of the proportion of variant sequences within a DNA sample.

UR - http://www.scopus.com/inward/record.url?scp=0033529765&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033529765&partnerID=8YFLogxK

U2 - 10.1073/pnas.96.16.9236

DO - 10.1073/pnas.96.16.9236

M3 - Article

VL - 96

SP - 9236

EP - 9241

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 16

ER -