Real-time PCR: housekeeping genes in the INS-1E beta-cell line

K Smidt; L Wogensen; B Brock; O Schmitz; J Rungby

doi:10.1055/s-2006-924968

Real-time PCR: housekeeping genes in the INS-1E beta-cell line

K Smidt, L Wogensen, B Brock, O Schmitz, J Rungby

7 Citations (Scopus)

Abstract

Investigation of gene expression is a developing area with several methods available. One method is quantitative PCR. A major pitfall in quantitative PCR is the normalisation procedure of the gene expression. Many experiments include a housekeeping gene, some use RNA concentration, and others use a geometric mean of several internal, stably expressed genes. This study demonstrates that real-time-PCR results differ with varying housekeeping genes and analysis protocols when applied to insulin-secreting INS-1E cells derived from the pancreas and stimulated by DEDTC (diethyldithiocarbamate, a zinc chelator) and GLP-1.

Original language	English
Journal	Hormone and Metabolic Research
Volume	38
Issue number	1
Pages (from-to)	8-11
Number of pages	4
ISSN	0018-5043
DOIs	https://doi.org/10.1055/s-2006-924968
Publication status	Published - Jan 2006
Externally published	Yes

Keywords

Animals
Cell Line
Chelating Agents
Ditiocarb
Gene Expression Profiling
Gene Expression Regulation
Humans
Insulin
Insulin-Secreting Cells
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Journal Article

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10.1055/s-2006-924968

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title = "Real-time PCR: housekeeping genes in the INS-1E beta-cell line",

abstract = "Investigation of gene expression is a developing area with several methods available. One method is quantitative PCR. A major pitfall in quantitative PCR is the normalisation procedure of the gene expression. Many experiments include a housekeeping gene, some use RNA concentration, and others use a geometric mean of several internal, stably expressed genes. This study demonstrates that real-time-PCR results differ with varying housekeeping genes and analysis protocols when applied to insulin-secreting INS-1E cells derived from the pancreas and stimulated by DEDTC (diethyldithiocarbamate, a zinc chelator) and GLP-1.",

keywords = "Animals, Cell Line, Chelating Agents, Ditiocarb, Gene Expression Profiling, Gene Expression Regulation, Humans, Insulin, Insulin-Secreting Cells, Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction, Journal Article",

author = "K Smidt and L Wogensen and B Brock and O Schmitz and J Rungby",

year = "2006",

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doi = "10.1055/s-2006-924968",

language = "English",

volume = "38",

pages = "8--11",

journal = "Hormone and Metabolic Research",

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TY - JOUR

T1 - Real-time PCR

T2 - housekeeping genes in the INS-1E beta-cell line

AU - Smidt, K

AU - Wogensen, L

AU - Brock, B

AU - Schmitz, O

AU - Rungby, J

PY - 2006/1

Y1 - 2006/1

N2 - Investigation of gene expression is a developing area with several methods available. One method is quantitative PCR. A major pitfall in quantitative PCR is the normalisation procedure of the gene expression. Many experiments include a housekeeping gene, some use RNA concentration, and others use a geometric mean of several internal, stably expressed genes. This study demonstrates that real-time-PCR results differ with varying housekeeping genes and analysis protocols when applied to insulin-secreting INS-1E cells derived from the pancreas and stimulated by DEDTC (diethyldithiocarbamate, a zinc chelator) and GLP-1.

AB - Investigation of gene expression is a developing area with several methods available. One method is quantitative PCR. A major pitfall in quantitative PCR is the normalisation procedure of the gene expression. Many experiments include a housekeeping gene, some use RNA concentration, and others use a geometric mean of several internal, stably expressed genes. This study demonstrates that real-time-PCR results differ with varying housekeeping genes and analysis protocols when applied to insulin-secreting INS-1E cells derived from the pancreas and stimulated by DEDTC (diethyldithiocarbamate, a zinc chelator) and GLP-1.

KW - Animals

KW - Cell Line

KW - Chelating Agents

KW - Ditiocarb

KW - Gene Expression Profiling

KW - Gene Expression Regulation

KW - Humans

KW - Insulin

KW - Insulin-Secreting Cells

KW - Reproducibility of Results

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Journal Article

UR - https://www.scopus.com/pages/publications/33645706276

U2 - 10.1055/s-2006-924968

DO - 10.1055/s-2006-924968

M3 - Journal article

C2 - 16477533

SN - 0018-5043

VL - 38

SP - 8

EP - 11

JO - Hormone and Metabolic Research

JF - Hormone and Metabolic Research

IS - 1

ER -

Real-time PCR: housekeeping genes in the INS-1E beta-cell line

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Keywords

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