Re: [DIYbio] Re: PCR trouble

I'd strongly recommend that you debug this with ampllifying a shorter fragment — something around 500 bp. You can do this quicker and easier than with the challenging long PCR you are attempting. Make a simple one work, then you will know where the problems arise.
How confident are you in the sequence of your template? Have you, for example, cut it with restriction enzymes and checked the length of fragments?

> On Mar 14, 2016, at 11:09 AM, David Ishee <midgardkennels@gmail.com> wrote:
>
> No product on either one, but also no unexpected product.
>
> It was probably my template, I was in a hurry and didn't use a miniprep, I tried to use a bit of the plasmid bearing bacteria. I'll try again with a fresh miniprep tonight.
>
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