E. Gottwald et al., Semiquantitative reverse transcription-polymerase chain reaction with the Agilent 2100 Bioanalyzer, ELECTROPHOR, 22(18), 2001, pp. 4016-4022
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.