Jump to main content
US EPA
United States Environmental Protection Agency
Search
Search
Main menu
Environmental Topics
Laws & Regulations
About EPA
Health & Environmental Research Online (HERO)
Contact Us
Print
Feedback
Export to File
Search:
This record has one attached file:
Add More Files
Attach File(s):
Display Name for File*:
Save
Citation
Tags
HERO ID
1235756
Reference Type
Journal Article
Title
Quantitative RT-PCR gene expression analysis of laser microdissected tissue samples
Author(s)
Erickson, HS; Albert, PS; Gillespie, JW; Rodriguez-Canales, J; Marston Linehan, W; Pinto, PA; Chuaqui, RF; Emmert-Buck, MR
Year
2009
Is Peer Reviewed?
1
Journal
Nature Protocols
ISSN:
1754-2189
EISSN:
1750-2799
Volume
4
Issue
6
Page Numbers
902-922
Language
English
PMID
19478806
DOI
10.1038/nprot.2009.61
Abstract
Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) is a valuable tool for measuring gene expression in biological samples. However, unique challenges are encountered when studies are performed on cells microdissected from tissues derived from animal models or the clinic, including specimen-related issues, variability of RNA template quality and quantity, and normalization. qRT-PCR using small amounts of mRNA derived from dissected cell populations requires adaptation of standard methods to allow meaningful comparisons across sample sets. The protocol described here presents the rationale, technical steps, normalization strategy and data analysis necessary to generate reliable gene expression measurements of transcripts from dissected samples. The entire protocol from tissue microdissection through qRT-PCR analysis requires approximately 16 h.
Tags
IRIS
•
Methanol (Non-Cancer)
Ramazzini
Other
•
Ramazzini Institute
Home
Learn about HERO
Using HERO
Search HERO
Projects in HERO
Risk Assessment
Transparency & Integrity