Real-time RT-PCR

Real-time RT-PCR is an established technique for quantifying mRNA in biological samples, this technique enables to carry out two different methods of quantification gene expression based on the porpouse of the study:
  • Absolute quantification:
    To obtain the quantity of nucleic acid (copy number, µg) per given amount of sample (per cell, per µg of total RNA)
  • Relative quantification:
    To obtain the relative amount of a target nucleic acid for equivalent amounts of test and control samples
This tool is designed for relative quantification analysis of expression pattern data measured by Real-time RT-PCR with ABI 7900 Sequence Detection (Applied Biosystems).
During analysis process algorithm follows some steps to obtain an optimum data analysis.
  1. Normalization against reference gene
  2. Parametric statistical analysis
  3. Graphical results

Here you could show an example of how to use the application presented next.

Data analysis
-Table of samples file to process:
-Raw qPCR data file:
-Enter title ;
-Enter housekeeping genes (separated by /)
ex:HPRT1-Hs99999909_m1/18S-Hs99999901_s1
-Enter Samplegroup id equivalence (separated by ;)
ex: 1,"controls";2,"malalts"
-Efficiency data file:
if suposed efficiency is equal to 2, no file are needed

Problems-Suggestions: [email protected] / [email protected]