we have got RT-PCR form applied biosystems, with out using fluorescent reporter probe in the reaction mix, can we use RTPCR for regular molecular biology work like amplification of genes,plasmids etc..
Hi Ravi-- I have often run endpoint PCR in a realtime instrument. Just make sure the melting and cycling parameters are appropriate; realtime experiments usually use short cycle times to amplify short sequences. Also, many realtime instruments cannot cool to 4 degrees at the end of the run; so don't leave your samples in the machine overnight before running your gels, etc.
Yes, you may run regular PCR amplification on real time PCR machine. The same process. Also, after validation of new primers by qPCR I always run PCR product on 2% gel ( second line of validation).
You can use it but it will not be cost effective for only a few samples if you are not able to use single tubes in the cycler. Make sure that you exclude imaging step from the program to save the precious lamp hours.
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