3 replies to this topic

[noyara]

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

Thank you

[lucky2610]

noyara, on 28 March 2011 - 08:07 PM, said:

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

Thank you

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)

[Nephrite]

noyara, on 28 March 2011 - 08:07 PM, said:

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

Thank you

Hi :-)

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

Check these two links:

http://www.bio.david...production.html
http://www.bio.david...dompriming.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

Edited by Nephrit, 29 March 2011 - 12:59 AM.

[noyara]

Thank you very much