I do Q-PCR using Taqman assays on the ABI-7500, I came across LinReg analysis tool for analysis!! But, when I export raw data into LinReg tool all data points are negative and an error message appears "Invalid floating point operation". Has anyone experienced this? Any help is greatly appreciated.
Regards,
Jeru Manoj Manuel
Have you checked that your "raw" data were not automatically baseline corrected by the software you used to record them? I experienced that getting true raw data from the software issued with the thermocycler and putting them in LinReg format could be quite a pain.
Hello Charlotte,
Thank you for the response.I did try that too, but as you have mentioned getting the true raw data seems an issue.
I directly exported from ABI 7500 system after removing the baseline correction. I am sorry for my non-awareness, is there any other software?
I haven't done the manipulation recently, but from what I remember, after exporting the raw data from SDS (probably same software for you as we were also running an ABI system), they were very strangely formatted with a few line per well and the various data for each time step in the different column. There were also empty lines and header lines between each well. So we had to reformat them manually to get a file with one column per type of measure, and additional columns indicating the well, the time step, etc... (see attached file, where I only copied the first lines of the 47777 total lines of the original file). That worked fine for us.
Kind of you for sharing a working format. The raw data formatting obtained was as mentioned by you. Now i have learnt how to go about arranging the data, thanks to your help.
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