oligo dt and random primer - (Mar/28/2011 )

Hi all,
What is the different between oligo dt and random primer? and what is the best for PCR?

Thank you

noyara on Tue Mar 29 04:07:01 2011 said:

It depends... If you work with prokaryota using oligo dt is nonsense... If you work with eukaryota, both ways could be suitable... I recommend you try it! Buy both types, run RT-PCR and then measure Cq values and you will see;o) For any gene could be result different...it depends on used transcriptases, temperature, secondary structure of target mRNA etc.

I also know that some companies offer the mix oligo dt and random primer...it could be also very interesting choice;o)

noyara on Tue Mar 29 04:07:01 2011 said:

Hi :-)

1. What is the difference between oligo dt and random primer?

Check these two links:

http://www.bio.davidson.edu/courses/genomics/method/cDNAproduction.html
http://www.bio.davidson.edu/courses/genomics/method/randompriming.html

They will explain you all about priming.

2. What is the best for PCR?

I think it depends on where your amplicon of interest is situated and how much your RNA is fragmented. I work with RNA from paraffin embedded sections. I have very good results using the random primers and very promising, when I tried gene-specific primers.

Nephrite

Thank you very much