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 1,340 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 1,037 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 775 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 4,059 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 7,903 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 4,157 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 501 0 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Why are Sanger Sequencing reads interrupted in some edited clones?

I have a Crispr/Cas9 edited bulk population and I can briefly check the indel ratios in the bulk population using Sanger sequencing. After cloning single cells, I wanted to check the indel types...

18 February 2021 8,198 6 View

Hi, can anybody help me with the suggestions how can I amplify a stress induced gene of 1107 bp?

I am trying to amplify the TGA1 (Transcription factor 1 ) of Arabidopsis ,which is 1107 bp. I have treated the plants with Salycylic solution over night and extracted the RNA and make cDNA. I...

17 February 2021 7,802 2 View

Alternatives to traditional cloning of insert into backbone (dsDNA)?

Hi, I am stuck with a cloning experiment: The insert & backbone(bb) share one compatible end at 3' end(NotI) and one incompatible end at 5' end (XbaI for insert; HindIII for backbone). So i...

16 February 2021 1,664 3 View

Why do I have trouble with cloning duplicate gene in the same plasmid?

Hello! I am trying to generate a construct in this format: seqA - seqB - linker - seqC - seqB into pSF-CMV-Ub-Puro-Ascl plasmid 0.5kb 2kb 66 0.25kb 2kb I have tried many...

15 February 2021 7,118 3 View

Would using an asRNA for repressing transcription of a high expression Listeria monocytogenes non-coding RNA harbored in a plasmid be a viable option?

Hello all, I am interested in deleting a highly expressed non-coding RNA located on a Listeria monocytogenes plasmid. Currently, there are not any optimized gene deletion methods for Listeria...

09 February 2021 4,994 1 View

Quality of antibodies from Creative Biolabs?

It seems Creative Biolabs has everything and so many versions of antibodies not owned or sold by any other company. How reliable are their products? I am looking for a clone where potentially...

08 February 2021 2,167 3 View