[Akira]

Hi. I have my own customized oligos and I would like to anneal them to make adapters. I've seen many protocols (including the Meyer's one) and I am confused whether to use Annealing Buffer with EDTA or without EDTA.

Now I have the oligos in lyophilized form: in which buffer should I resuspend them? H2O or TE or Tris-HCl?

And for Annealing Buffer:
Tris+ NaCl + EDTA? or TrisHCl only?

I will use NEBNext kit to do the ligation. Will EDTA interfere with the ligation process?

Please help. =(

[joze]

Hello everyone,

I realise this thread is pretty old now, but would anyone be able to attach the Meyer paper? I can't access it without subscription.

Cheers,

Jo

[Aska]

Hi there,

I was wondering if anyone could let me know what taq polymerase have they used for the multiplex PCR prior to HiSeq Illumina sequencing? Or what are they ways to chose one

[joze]

Hello there,

I was advised to use a High-Fidelity polymerase (I used the Phusion High-Fidelity polymerase). Note that the reaction conditions are slightly different using these.

Good luck,

Joze