Sonalika Mahajan

10 Questions 20 Answers 0 Followers

Questions related from Sonalika Mahajan

Can somebody please provide suggestions as I am struggling to express ACAP2 protein in mammalian cells?

I have cloned ACAP2 in pCMV-HA vector and the frame has been sequence confirmed. I am trying to express in HEla cells by transfecting he plasmid at 2.5µg per well (6-well plate) using...

09 September 2015 6,450 3 View

How can I express GST-tagged recombinant protein in soluble form and prevent the inclusion body formation?

I have expressed GST-tagged protein in BL21(DE3)pLysS cells but I am not able to get the protein in soluble form. Most of the protein is in inclusion body. The induction conditions used are 1mM...

05 May 2015 9,265 9 View

What is the best method to lyse bovine kidney cell line?

I want to perform Co-IP from the bovine kidney cell line transfected with the Myc-tagged bait and HA-tagged prey proteins. I am using Pierce RIPA lysis buffer for the cell lysis but the lysate I'm...

03 March 2015 2,810 7 View

Where do I start screening for RNA-protein interactions?

I want to identify the host proteins that interact with the non-coding RNA of virus. Is it possible to do the screening on the similar lines as done in protein-protein interactions using Y2H.

02 February 2015 357 5 View

How to calcuate the dose of virus needed to infect 25cc flask at 1000 TCID50?

I have estimated the virus titre to be 10 6.5 TCID50/50 µl. I want to know that how much virus I needed to add to infect 25cc flask at 1000 TCID50?

11 November 2014 5,488 3 View

Can somebody please suggest some tools to perform Gene ontology t0 the third level using UniProt IDs of the genes ?

I have screened a cDNA library of swine. I want to perform gene ontology on the obtained gene sequences so that a pie diagram representinbg the proportion of genes participating in various...

10 October 2014 547 11 View

Can anyone lend advice on CDNA library construction?

I am trying to construct a cell line library for yeast teo hybrid mating. the mRNa concentration was low so I concentrated using a vacuum concentrator and the final conc. of mRNA was 370 ng/µl. I...

07 July 2014 8,250 4 View

Has anyone expressed Nus tagged fusion protein?

I am trying to express one protein in pET-43.1 vector fused with N-terminus Nus tag but the size of the expressed protein is about 20 kDa smaller than expected. I also transformed the neat...

04 April 2014 1,388 1 View

Can somebody suggest any alternate method of Acetone protein concentration and analysis?

I have expressed one viral gene in Pischia pastoris. the expressed recombinant protein is secreted in the supernatant. I have concentratexd the supernatant using acetone and TCA method and run the...

04 April 2014 2,192 3 View

how to make a 5-25% gradient SDS-PAGE?

I want to run rAB on gradient SDS-PAGE. I tried twice but the gradient was not formed. Can somebody please send me the protocol?

09 September 2012 7,437 2 View