English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Housekeeping gene selection for quantitative real-time PCR assays in the seagrass Zostera marina subjected to heat stress

Ransbotyn, V., & Reusch, T. B. H. (2006). Housekeeping gene selection for quantitative real-time PCR assays in the seagrass Zostera marina subjected to heat stress. Limnology and Oceanography: Methods, 4, 367-373.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000F-D922-C Version Permalink: http://hdl.handle.net/11858/00-001M-0000-000F-D923-A
Genre: Journal Article

Files

show Files
hide Files
:
Ransbotyn_V_Reusch_TBH2006.pdf (Publisher version), 85KB
 
File Permalink:
-
Name:
Ransbotyn_V_Reusch_TBH2006.pdf
Description:
-
Visibility:
Restricted (Max Planck Institute for Evolutionary Biology, Plön; )
MIME-Type / Checksum:
application/pdf
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show

Creators

show
hide
 Creators:
Ransbotyn, Vanessa1, Author              
Reusch, Thorsten B. H.1, 2, Author              
Affiliations:
1Department Ecophysiology, Max Planck Institute for Limnology, Max Planck Institute for Evolutionary Biology, Max Planck Society, ou_976547              
2Department Evolutionary Ecology, Max Planck Institute for Evolutionary Biology, Max Planck Society, ou_1445634              

Content

show
hide
Free keywords: -
 Abstract: In light of increasing sea surface temperatures, quantifying the expression of stress-inducible genes in coastal organisms is an important topic of marine molecular ecology and evolution. As a prerequisite for quantitative realtime PCR (QPCR) assays, we tested the suitability of 12 candidate housekeeping genes (HKGs) for normalization of messenger RNA abundance and quality in the ecologically important seagrass species Zostera marina. In comparing several algorithms used to identify appropriate HKGs, two approaches using reciprocal cross-validation among a larger group of HKG candidates implemented in geNorm or NormFinder yielded largely consistent results. Using these approaches, 3 HKG candidates were selected that are stable alone or in combination in the target tissue (leaf) with respect to the imposed temperature stress treatments (TATA box binding protein, translation elongation factor, eukaryotic initiation factor 4A). Example calculations revealed spurious gene expression changes of a factor of 2 in hypothetical target genes when using less stable HKGs as reference for standardization. These results highlight the need for devoting considerable effort to selecting appropriate HKGs for normalization of QPCR data before performing experiments with the target genes.

Details

show
hide
Language(s): eng - English
 Dates: 2006
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: eDoc: 289991
Other: 2490/S 38531
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Limnology and Oceanography: Methods
  Alternative Title : Limnol. Oceanogr.: Methods
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 4 Sequence Number: - Start / End Page: 367 - 373 Identifier: ISSN: 1541-5856