1 reply to this topic

[izzybusydizzy]

Good day everyone, I can use some advice from you.

I am amplifying a new target and have not choice as to the placement of my primers (start of the gene and end of the gene)

The primers have a high affinity to prime to themselves as well as each other.Below are the specs of my primer pair.

I have not worked with primers that are this bad. I'd appreciate any tips from anyone who have worked with similar primers.



Length (bp) GC% Tm Hairpin   Self-Dimer Hetero Dimer
21        47.6 57.6 -0.69   -9.28         -14.77
20        55 58.2 -2.03   -9.28         -14.77


Thank you everyone.

[Nephrite]

izzybusydizzy, on 17 March 2011 - 09:46 AM, said:

Good day everyone, I can use some advice from you.

I am amplifying a new target and have not choice as to the placement of my primers (start of the gene and end of the gene)

The primers have a high affinity to prime to themselves as well as each other.Below are the specs of my primer pair.

I have not worked with primers that are this bad. I'd appreciate any tips from anyone who have worked with similar primers.



Length (bp) GC% Tm Hairpin   Self-Dimer Hetero Dimer
21        47.6 57.6 -0.69   -9.28         -14.77
20        55 58.2 -2.03   -9.28         -14.77


Thank you everyone.

Hello.

I have the same problem but can`t help. There is a pinned topic ''How to get rid of PCR primer dimer'' - may be you should go there and have a look. Probably you will find some decision.

Good luck.

Nephrit.

Edited by Nephrit, 18 March 2011 - 01:27 AM.