Jorge Versicolor

6 Questions 2 Answers 0 Followers

Questions related from Jorge Versicolor

Does DNA A-tailing survive after agarose gel electrophoresis and purification?

Hi everyone, I need to clone in pGEM-T vector the result of amplifying a metagenomic library by PCR with a high fidelity polymerase but I only want to clone the fragments of an specific size...

05 February 2021 6,280 8 View

How can I solve this problem with agarose gel DNA purification of bigger (1-4kb) fragments ?

Hi everyone, I perform a PCR + Restriction Digestion (single cut, same at both sides) over a metagenomic library. After that, I am trying to purifiy DNA fragments between 1-4kbp long from the...

25 January 2021 2,437 5 View

Is there any High-Fidelity DNA polymerase able to work in droplet (digitalPCR)?

Hi everyone I need to amplify DNA inside droplets made with a microfluidic device. I tried Taq based polymerases and yield is good but error rate is higher than what I need. I also tried with...

27 February 2020 9,612 0 View

How to reduce droplet merging in droplet digital PCR (dPCR) ?

Hi everyone. I am performing PCR in droplets of 20um of diamater and my protocol is a classical PCR protocol, oil HFE 7500 and surfactant is PicoSurf 3%. I leave like 20uL of oil on the bottom of...

30 October 2019 3,048 3 View

Problems to construct a plasmid library with long metagenomic fragments. Any help?

Hello everyone! I am trying to make a library of metagenomic DNA in the plasmid PBlueScript SK+. I fragmented metagenomic DNA by sonication and I confirmed the average size of the fragments is...

09 August 2019 7,359 5 View

How to avoid self ligation of large inserts during clonning?

Hello everyone, I've been trying to perform a ligation for the last couple of weeks and I just get empty plasmids. The insert is pool of fragments amplified by PCR with an average length of 3kb...

15 April 2019 9,765 5 View