Juan Pablo Morán

7 Questions 4 Answers 0 Followers

Questions related from Juan Pablo Morán

Can a 5 nt gRNA spacer perform like a 8 nt-cutter restriction enzyme using CRISPR-Cas system?

Or do you know what is the inferior limit in spacer length? 5+3 from the PAM=8

10 November 2021 5,132 2 View

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 5,032 3 View

Are Aspergillus flavus and Aspergillus oryzae similar in their hydrolytic capabilities?

Aspergillus flavus and Aspergillus oryzae are very closely related species (just as A. parasiticus is with respect to A. sojae). However, A. flavus is pathogenic and A. oryzae is GRAS. It has...

21 September 2018 7,428 4 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 4,784 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 6,332 4 View

A particular E. coli strain, non deficent in homologous recombination enzymes, should be used for PCR fusion cloning?

We are doing modifications of an expression vector -a promoter substitution-, and I was wondering if attempting this task through PCR cloning using overlaped homologous sequences could be done by...

04 May 2017 7,643 1 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 4,121 5 View