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

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

Short Synthesis of Graphene Oxide from Natural Graphite Flakes?

I am in search of a modified Hummer's method, which can be used to synthesis graphene oxide within 6-8 hours. As I am a student it is not allowed for us to work after 5 in the laboratory. So I am...

01 August 2024 8,368 2 View

Some new emerging problems on application of RL for scheduling in IoT networks?

I have seen plenty of existing works on applied Reinforcement Learning (RL) policies for optimized scheduling in IoT networks including Q-learning, DQNs, and the newer ones including PPO for...

01 August 2024 8,754 2 View

How to make soil water characteristic curve using centrifuge?

We have no access of pressure plate apparatus

22 July 2024 3,488 2 View

What is the degassing temperature and time for biochar for BET surface area analysis if TGA is not possible?

Since BET and TGA both are very costly instruments and many places the facilities are not available (for large number of samples) hence a threshold degassing temperature and time is preferable

16 July 2024 8,112 2 View

Can we analyse RMSD for peptide-HLA docking and binding affinity for peptide-HLA to TCR docking, in a peptide-HLA-TCR docking ?

In the peptide-HLA-TCR docking process, can we express the result of peptide-HLA docking in RMSD and for peptide-HLA to TCR docking, can results be expressed in binding affinity?

11 July 2024 4,831 0 View

How to get discharge capacity of a bole well?

No data regarding radius of influence is provided and no pumping test data available.

10 July 2024 7,168 0 View

How to apply an external electric field in gaussian 16 or Material studio DMOL3 ?

I need to apply an external electric field to 4-hydroxybenzoic acid to compare the molecule's reactivity descriptors. Could you guide me on how to apply the electric field using Gaussian and...

01 July 2024 4,192 3 View

How can I precipitate a protein from condition media ?

So I want to see is the level of a secreted protein in the condition media via western blotting What my doubt is 1. How much cells should I seed and what should be the volume of condition...

19 June 2024 1,927 1 View

Ni-NTA protein purification - how to obtain higher yield concentration?

Hello, I am trying to purify a 6X-His tagged protein using Ni-NTA chromatography. However, I obtain very less yield & concentration; and considerable protein in load flow through. Any...

16 June 2024 5,876 5 View

In sds page and western blot, non specific expressed protein with very thin band is also there along with thick band of specific protein. why ?

Asper discussion, First time my clonning was failed becuase the first 45 nucleotide was not cloned in pet30 a vector then I choosed the different clone of 1:1 ration fraction, I got the size of...

13 June 2024 7,564 7 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