In Gibson cloning, how many base pairs does 5' exonuclease chew up?

More Sanjan T.P. Gupta's questions See All

What are the most reliable approaches to extract key continuous frames from a video, with complete automation?

I have an input video, of skin (from a microscope that means it is magnified such that blood capillaries are visible). The idea is to extract 15 sec of continuous frames from a video of 1 min,...

02 March 2021 2,735 2 View

Which microtomy technique do you suggest for thin and delicate root sectioning ?And, how to preserve and process the roots before sectioning?

We have thin roots of from 1 year old different species of plants. We are looking into anatomical details of the roots. What's the best method to preserve and process the roots once they are...

23 February 2021 1,898 1 View

Has anyone done Archard Wear Simulation of GFRP Composite Plate in ANSYS with ACT extension or UDF?

I'm looking to do a sliding wear (plate-on-plate) simulation of GFRP composite plates with Archard Wear Model. Any hints or example steps are appreciated.

17 February 2021 1,447 2 View

Which root in this tap root system, in the picture attached, should be used for root sectioning in order to study the root anatomical details?

There are numerous roots in this tap root system. Kindly suggest or share a research paper where I can clarify the doubts as to which root should be chosen for sectioning and for further root...

14 February 2021 1,816 3 View

Possible problem for showing 0.2 sun intensity instead of 1 sun intensity in AAA class Newport solar simulator?

We have a AAA class Newport solar simulator. We changed the Xenon Arc Lamp recently (XBO450W, Make:OSRAM) as the lifetime got over. However, the intensity is coming to be only 0.2 sun instead of 1...

14 February 2021 9,998 3 View

How can I get access of UNAfold tool?

I am searching for tools to construct molecular beacons which shall be complementary to a given sequence. I don't want to design any primers. I found that UNAfold is one such tool which I could...

02 February 2021 9,065 5 View

The peptide synthesized with CTC resin have mass more (+68 Da) than the desired mass of the synthesized peptide. What could be the reason?

I am synthesizing 7 amino acid long peptide sequence with CTC resin, 50oC coupling for 10 min. On cleavage (TFA/TIPS/Water 95:2.5:2.5) I got peptide with good yield. However, It has 68 Da more...

30 January 2021 8,483 3 View

How do we calculate curvature value in ArcGIS?

Hello everyone, I am trying to prepare curvature map in ArcGIS 10.4, using spatial analyst tool. I am using 10 m CartoDem (projected in UTM 44N). I am getting very different value, curvature...

12 January 2021 504 4 View

How can a publish my case report?

I want to publish a case report regarding jejunal diverticula

02 January 2021 7,630 5 View

How can we maintain the two different DC high voltages of same polarity between parallel electrodes inside the vacuum separated by few millimeter?

I am trying to maintain two different high DC voltages of same polarity (either positive or negative) between two parallel electrodes separated by 500 um inside the vacuum. However, what I found...

21 December 2020 1,735 3 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