Can anyone suggest how to clone a gene of lignin peroxidase in DHa 5 E.coli?

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What essential parameters should be checked for primer designing, and what software should be used?

My present research is on primer designing for lignin peroxidase and laccase gene.

10 November 2013 5,687 8 View

What is the best method to prepare the competent cells of E.coli for cloning?

what will be the best method to prepare the competent cells of E.coli for cloning?

08 September 2013 4,061 5 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

How can we now if the promotor in the vector is well?

Hi, everyone. I want to transfer two genes in to the pseudomonas bacteria that isolated from the soil. The bacteria that I want to use for cloning haven't been identified and not be sequenced,...

02 August 2024 3,987 1 View

Illustra™ MicroSpin™ G-25 columns what it is used for?

Hello, We found three packages of Illustra™ MicroSpin™ G-25 columns in the cabinet of an unused lab. They are very old but have never been opened. I have never used this kit before, and I couldn't...

25 July 2024 4,927 3 View

Pink bacterial colonies?

I am cloning an overexpression plasmid with my protein of interest tagged with mScarlet. After transforming my ligated product into DH5α bacteria and plating on LB agar, I noticed colonies with a...

22 July 2024 5,953 6 View

What are the strategies to Enhance IgG-Producing Plasma Cells in mice for Monoclonal Antibody Development?

I have immunized BalB/C mice with a protein using the intradermal (ID) method with Complete Freund's Adjuvant (CFA) and Incomplete Freund's Adjuvant (IFA), following a 14-day interval and three...

22 July 2024 9,160 2 View

Cloning of flaviviruses is difficult i have noticed that cells loose their transformation ability after cloning fragments inside vector any advice?

As flaviviruses genome is very frequently prone to mutation hence it is very difficult to clone whole genome inside vector what kind of vector and cells can sustain such amount of cloning...

11 July 2024 4,284 0 View

I am not getting proper colonies while ligating one of fragments in my vector the colonies seems to be normal but in light it seems lysed any reason?

I am working on cloning of a gene fragment inside my vector everything we tried to do correctly like making insert and vector with utmost care but after ligation the morphology of colonies seems...

11 July 2024 9,010 3 View

How can I verify if an academic journal is a legitimate publication or a cloned version?

This question seeks to address the growing concern of cloned academic journals, which are fraudulent duplicates of legitimate publications. It aims to guide researchers, scholars, and academics on...

10 July 2024 5,966 2 View

ATG in pPICZalfphaA vector: Yes or Not in front of the gene of interest ... ?

Is it appropriate to place the ATG codon in front of the gene of interest, since there is a secretion signal that has its own ATG in front of this gene? I need my protein of interest to be...

10 July 2024 3,050 5 View

I need help with cloning a 200 bp insert and a 4,7 Kb pEGFP-N1 vector. How can I do it?

I've been trying to clone my N-terminal inserts in the comercial pEGFP-N1 vector. Initially, I cloned my N-terminal insert into a pGEMT-easy vector to ensure that the insert digestion was done...

09 July 2024 4,277 1 View