Juan Pablo Morán

4 Questions 3 Answers 0 Followers

Questions related from Juan Pablo Morán

Does site-directed mutagenesis by subcloning works for you?

Hi folks! I want to subclone the S. pyogenes nickase (D10A) gene into certain expression vector by PCR-amplifying it from a Cas9 template and assemble this with a regular isothermal enzymatic...

17 March 2020 4,920 3 View

Do you know what can be the cause for this uniform all-lane smearing?

We are linearizing a vector with a HF polymerase (Q5 from NEB and its recommended parameters). I established conditions that produced a sharp and unique amplicon, but I've just tried it one more...

18 June 2017 4,752 4 View

Do you know the reason for a band in an agarose gel to look like the one at the right in the photograph?Is it a reaction or an electrophoresis issue?

Electrophoresis conditions: 1% agarose, TAE buffer, constant voltage at 7 V/cm electric field strength, and 10 ul were loaded. The amplicon comes from a touch down PCR.

19 May 2017 6,285 4 View

Does in vivo ligation requieres phosphorylated ends?

We will perform a simple cloning technique based on PCR. We have our covalently closed circular vector and an amplicon produced by asymmetric PCR that will serve as a megaprimer. Hence, it will...

27 December 2016 4,074 5 View