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

More Mahesh Agarwal's questions See All

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...

04 May 2018 2,075 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?

05 June 2016 7,250 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?

05 June 2016 6,170 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...

04 May 2016 6,608 8 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...

04 May 2014 3,476 0 View

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

03 April 2014 3,528 13 View

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

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

09 October 2012 1,042 9 View

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

Method used is ^^CT method

09 October 2012 1,451 23 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...

02 March 2012 8,472 13 View

Cytotoxicity activity of plant extract

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

08 September 2011 1,712 7 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

16 June 2021 0 0 View

fatal error

16 June 2021 0 0 View

fatal error

16 June 2021 0 0 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

What kind of fluid could I use for a pitfall trap that also does not invalidate molecular testing?

Hi, I want to start testing pitfall trap to obtain ants samples, but I need to conduct molecular analysis on those insects. So, what kind of fluid can I use? Ethanol expires too early and I need...

03 March 2021 5,978 5 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

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View