Ravi Shankar Gautam

8 Questions 23 Answers 0 Followers

Questions related from Ravi Shankar Gautam

Why I amGetting non specific bands for my MBP fusion protein after GPC?

Hello Every1!!! I am trying to purify my MBP fused protein by affinity chromatography using amylose resins but getting to many non specific bands.. therefore i tried gel filtration but again and...

08 August 2019 2,454 4 View

Double digestion problem ?

In order to do a cloning I am trying to do double digestion of my vector with Sph1 and Not1 enzymes , the sites are 14 bases apart. My question is - is it appropriate to do double digestion at a...

04 April 2019 627 3 View

How much concentration of template(Rec Plasmid) required to see a fallout of 166bp?

Hi all, I m very much confused for a double digestion reaction to check a fallout of 166 bp which has been cloned in 6900bp vector. I have took around 1500 ng of my recombinant plasmid(6900+166...

04 April 2019 7,058 4 View

How can I get Chromosomal location of fungal genes?

Hello all, I am looking for bioinformatic tool/websites to know the location of fungal genes, as most of the genes are uncharacterized , is it possible to get their location on chromosome? My...

03 March 2019 3,434 2 View

Any suggestion why i am getting higher protein band size??

Hello all! I am doing SDS-PAGE for my protein of predicted size of 53.5kda bt unexpectedly I am getting bands of around 60kda.. I have done this 3-4 times and Everytime the required band is about...

02 February 2019 5,619 8 View

Does Mutation In Ribosomal Binding Site cause any interference in protein Expression?

Does single nucleotide mutation in Ribosomal Binding Site of an expression vector hamper protein expression? As there is no band could be seen in SDS-PAGE..

08 August 2018 4,135 5 View

My clone ( from cDNA) after sequencing reveals 2bp deletion in , should I use SDM for repair?

My clone ( cDNA Clone) has 2 bp (C&A) deletion as a result their is frame shift and no protein expression is taking place. gDNA clone reveals no deletion but all my cDNA clone reveals 2bp...

07 July 2018 10,055 9 View

I m trying to clone one gene from cDNA(502bp) and gDNA(600bp), during PCR i m getting amplification from gDNA only not from cDNA, any solution??

Same cDNA is used for other amplifications and i m getting result for that ,, actually the Tm of the primers is too low unfortunately it is around 35 degree , when i put an PCR at Tm 28 i got some...

02 February 2018 6,761 6 View