Rohit Kunar

11 Questions 25 Answers 0 Followers

Questions related from Rohit Kunar

Can one isolate proteins from cell lysates using antibody and use the protein for structural bilogy experiments?

I have been trying to over-express my protein in bacteria but in vain. Alternatively, is it possible to isolate the protein from cell lines/tissues using an antibody and then purify the protein...

01 January 2019 1,712 5 View

Can anyone help me with the identification of Drosophila larval neurons shown here?

Dear Colleagues, I have been trying hard to identify the exact neuronal identity of the neurons shown in the image. It would help me greatly if anyone would help me out with their identification....

09 September 2018 6,416 0 View

In silico softwares for Prediction of Protein-RNA binding?

I am working on an RNA binding protein, against which I do not have an antibody. Neither is the crystal structure known in the particular organism. I am interested to have an in silico prediction...

06 June 2018 9,215 4 View

Can anyone help me with the Optimisation of Induction for a GST fusion protein?

I have a clone of my protein coding ORF in pGEX-4T-1 vector which should produce a fusion protein of ~115kDa. Snapgene Analysis of the clone (sequence) shows that the clone is in proper...

02 February 2018 4,587 7 View

Can anyone help me with IPTG induction in pGEX-4T-1 clone?

I induced two cultures at 0.5 OD600: 1 with just the vector, and another with my ORF in the vector. However, following induction with 1mM IPTG with shaking at 140 rpm at 37 degree C for even 6...

10 October 2017 5,556 5 View

Can gypsy elements hop in presence of P-transposase?

I have discovered a putative insertion at a certain locus in a mutant Drosophila. The mutation was actually a second site mutation in a P-element hop line. Primary end sequencing of the same...

01 January 2015 8,854 4 View

How can I get good Southern Blots with DIG Labelled probes?

I am trying to confirm the presence of a putative 8.5 kb insertion in a Drosophila mutant, by PCR and/or Southern Blotting. I use almost 10 ug DNA for each lane, and specific probes(812 bp) which...

12 December 2014 276 2 View

How can I clone a PCR generated 9 kb amplicon in a 7.8kb vector?

I am trying to clone a full length gene with flanking sequences (a total size of 9 kb) in the Drosophila expression vector pCaSpeR4. I had constructed primers with sites for BamHI and KpnI in the...

12 December 2014 7,182 13 View

Are riboprobes better than DNA probes for southern?

I have been trying Southern with DNA probes (DIG labeled), but my blots are unsuccessful. Is it worth trying with riboprobes, dig labeled again? Can someone provide a protocol which one has...

12 December 2014 6,946 3 View

How to confirm putative long novel insertion in genome by PCR?

I am trying to locate a putative insertion of almost 8.5 kb in a Drosophila mutant by Long Range PCR using Platinum High Fidelity Taq Polymerase. The insertion does not appear in every PCR,...

12 December 2014 1,318 4 View

Are AT-rich long insertions in genome, omitted in NGS?

I have a Drosophila mutant, wherein I suspected the presence of an 8.5 kb insertion in the genome, by PCR. End sequencing of the same revealed an AT-rich sequence. To confirm its presence and in...

12 December 2014 3,088 6 View