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Primers and self and hetero dimers


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#1 izzybusydizzy

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Posted 17 March 2011 - 09:46 AM

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.

#2 Nephrite

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Posted 18 March 2011 - 01:23 AM

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.





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