Mahesh Agarwal

11 Questions 8 Answers 0 Followers

Questions related from Mahesh Agarwal

Why I am getting multiple band in SDS-PAGE after protein purification?

I have cloned a gene (750 bp) in pMAL-p5x expression vector. My protein is of size 29 kDa and it is fused with Maltose binding protein (42 kDa). I got 3 bands in SDS-PAGE after the protein...

05 May 2018 2,140 3 View

Agarose gel percentage to see band of 8 kb and 0.1 kb?

I have double digested a vector having size 8 Kb and Insert 100 bp (0.1 Kb). What percent Agarose gel I used use to see both band clearly?

06 June 2016 7,308 16 View

Which E.coli Strain is best for expression of protein from MBP vector?

Can anybody send me detail protocol for expression and purification of protein from MBP vector?

06 June 2016 6,213 4 View

What is the possible reason for getting smear in a gel?

I found smear in gel after double digestion of Plasmid with SfiI and NotI (Lane 4). Single digestion with SfiI works fine (Lane 3). Lane 2 is uncut plasmid. What is the possible reason for getting...

05 May 2016 6,667 8 View

How can I perform gene knock in and knock out simultaneously in a cell line?

I want to insert a GFP gene in a particular locus of a chromosome by replacing already present gene at that site. Which approach will be highly efficient for homologous recombination? Which vector...

05 May 2014 2,129 2 View

How to insert GFP gene in a genomic DNA of U-87 MG (glioblastome cell line)?

Is it possible to insert a GFP gene in gDNA of U87MG by replacing some other gene at that place? If, yes then which GFP vector should I use to transfect in U87MG? Kindly tell me detail procedure...

05 May 2014 3,526 0 View

How can I estimate the copy number of a gene by using SyBR green realtime PCR?

04 April 2014 3,584 13 View

Which is the best freely available software for quantifying cDNA by using Real time PCR?

Method used is ^^CT method

10 October 2012 1,518 23 View

Which is the basic book for understanding Fluorescence-activated cell sorting ?

Especially, the instrument set-up and Data analysis????

10 October 2012 1,082 9 View

How to reduce star activity in restriction digestion?

I am using BamH1 and EcoR1 for restriction digestion of my gene of 1.3Kb but EcoR1 is also cutting in between the gene. I am getting two fragments of 900bp and 400bp.This is due to presence of...

03 March 2012 8,527 13 View

Cytotoxicity activity of plant extract

Which is the standard cell lines used for checking cytotoxicity activity of medicinal plant extract ?

09 September 2011 1,768 7 View