How can I overcome binary vector ligation troubleshooting?

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npt II gene is possible to detect siRNA in plants.

I got the result from those region. If possible?

07 August 2013 1,433 0 View

How to check whether T4 ligase enzyme is working?

I am using fermentas T4 DNA ligase enzyme for ligate binary vector with 2.7kb insert. But it does not show any positive colonies. How can I check that enzymes is working?.

07 August 2012 1,140 2 View

Problem with binary vector cloning. Is there any method available to check for our cloning product?

I have ligated 1.3kb fragments to binary vector but it is not showing any single colony after transformation. What could be the problem?

07 August 2012 869 7 View

Could anybody help me with PCR troubleshooting?

I have done PCR many of the times with the same DNA, sometimes I got good results, sometimes they did not show any specification. What is the problem for this condition or do I need to change any...

06 July 2012 4,241 12 View

How to select the sequence for designing siRNA?

I want to know about primer designing for RNA interference. Which region should be selected for efficient target degradation? I am working with ssDNA geminviruses.

06 July 2012 7,984 0 View

Restriction enzymes digestion for release of insert from vector?

I want to know about the release of an insert from a vector. I am using two restriction enzymes for release of insert. It binds with a vector containing restriction sites. How can I release that one?

06 July 2012 4,265 3 View

Binary vector cloning

I want to know about binary vector cloning, because many binary vectors are using to our research purpose. Whether it is possible to know binary vector construction?

06 July 2012 588 0 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