Linear dynamic range for signal detection in fluorescent differential display.
We have determined the linear dynamic range in signal detection by Fluorescent Differential Display (FDD) using conditionally induced mRNA expression of the p53 tumor-suppressor gene as a control. By serial spiking of p53-induced RNA into that of non-induced RNA, we were able to quantitatively measure up to 100-fold change in p53 mRNA expression level. The linear dynamic range of signal detection per mRNA message was determined to be from 1000 up to 20,000 in fluorescence signal, in which the signals for the majority of mRNAs reside. Thus, FDD can be used to accurately quantify differences in mRNA expression among eukaryotic cells.
Cho
Yong-jig
YJ
Department of Cancer Biology, Vanderbilt-Ingram Cancer Center, Vanderbilt University Medical Center, 546 PRB, Nashville, TN 37232, USA.
Meade
Jonathan D
JD
Shester
Blake R
BR
Walden
Jamie C
JC
Guo
Zhen
Z
Liang
Peng
P
D005453:Fluorescence; D020869:Gene Expression Profiling / Q000379:methods*; D016158:Genes, p53; D012333:RNA, Messenger / Q000096:biosynthesis; D012680:Sensitivity and Specificity
D012333:RNA, Messenger