Semiquantitative reverse transcription-polymerase chain reaction with the Agilent 2100 Bioanalyzer

Citation
E. Gottwald et al., Semiquantitative reverse transcription-polymerase chain reaction with the Agilent 2100 Bioanalyzer, ELECTROPHOR, 22(18), 2001, pp. 4016-4022
Citations number
17
Categorie Soggetti
Chemistry & Analysis
Journal title
ELECTROPHORESIS
ISSN journal
01730835 → ACNP
Volume
22
Issue
18
Year of publication
2001
Pages
4016 - 4022
Database
ISI
SICI code
0173-0835(200110)22:18<4016:SRTCRW>2.0.ZU;2-Z
Abstract
We have applied a method to monitor mRNA expression in a semiquantitative f ashion on the Agilent 2100 Bioanalyzer. The method was originally described in 1994 by Wong, et al. and referred to as the ,,primer-dropping" method. This polymerase chain. reaction (PCR) technique uses multiple sets of prime r pairs in a coamplification reaction that amplifies the target of interest within a predetermined range specific for each target. Separation, detecti on and quantification of PCR products were accomplished using the Agilent 2 100 Bioanalyzer in conjunction with the DNA 500 and the DNA 1000 Lab-Chip k its for the detection of DNA fragments with a maximum size of 500 and 1000 bp, respectively. Using primers specific for the inducible form of hsp72 an d primers for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an intern al standard we were. able to rapidly monitor and quantify inducible hsp72-m RNA expression.