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Troubles with Fusion PCR

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Posted 28 March 2014 - 11:41 AM

I am trying to make a fusion PCR product with a 5.5kb and a 720bp band. However, I am not getting anything after I anneal and elongate the products. I am using equimolar concentrations of both DNA products and I have an extension time of 6.5 min at 68C. Does anyone have any experience with fusing large fragments that could give me some pointers? I have tried adding more dNTPs and more Taq polymerase to the mixes, but that has not worked for me. 

#2 bob1


    Thelymitra pulchella

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Posted 02 April 2014 - 12:05 PM

Taq will struggle with generating products of that size, try a long read polymerase such as KOD or phusion.

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