David A Johnson

5 Questions 9 Answers 0 Followers

Questions related from David A Johnson

Can I use a Taq mastermix for A-tailing?

I have gel-purified PCR product from a Q5 high-fidelity amplification to which I want to add the A tails for TOPO-TA cloning. The protocols I find all say to use Taq polymerase and dATP, but can I...

09 September 2019 6,865 3 View

For inverse PCR, is it OK if my restriction enzyme cleaves my known DNA once?

I am running inverse PCR to try to sequence the region flanking a novel gene. My question is whether or not it is OK to use a restriction enzyme that cleaves my known sequence at one site. That...

07 July 2019 5,462 4 View

How do I design inverse PCR primers for environmental sample with multiple sequences?

I have an environmental genomic DNA sample that includes three different newly-discovered 134 nt novel sequences. I want to do inverse PCR (using restriction enzymes and ligation) to begin...

05 May 2016 4,199 1 View

Can I use PCR reaction run with normal primers as template source for inverse PCR reaction?

I will be running inverse PCR to sequence the segments adjacent to my target segment. One of the template sources will be very difficult obtain again. I first ran the non-inverse PCR to discover...

04 April 2016 2,133 4 View

Paramecia in situ PCR?

I am attempting in situ PCR with paramecium. Has anyone tried in situ PCR in a similar organism with a pellicle? I trying to determine the parameters for fixation and protease treatment. Any help...

11 November 2015 3,019 1 View