Subramaniyam Ravichandran

12 Questions 33 Answers 0 Followers

Questions related from Subramaniyam Ravichandran

Band broadening in sequencing gels: How to avoid?

Hello all, I have been running sequencing gels for quite some time now and I find that the bands start broadening as the dye reaches the halfway mark (pic attached). I followed the publications...

04 November 2018 4,264 8 View

Do Protein A/G agarose beads nonspecifically bind to DNA?

Hi all, I am performing a Co-IP experiment where would like to check the binding of my protein to the DNA of interest. I kept a control where I incubated the DNA and antibody only with the...

09 May 2018 6,454 5 View

How can I calculate the concentration of protein in the enzyme from NEB?

I would like to calculate the concentration of T7 polymerase in the enzyme from NEB. Can anyone tell me how I can do this? Thanks in advance.

22 April 2018 6,365 3 View

Is it advisable to directly compare read counts for differential expression analysis?

I have an experimental setup where the RNAseq reads have been quantified using Htseq-count and then the DE analysis was performed using GFOLD software. In the original count files of control vs....

17 November 2016 1,750 4 View

Open Science Framework is a great tool for project management and collaboration, but is there a similar system that relies on local storage instead?

I have used OSF and love it, but my data has now overgrown the limitations of Google Drive and Dropbox. Since I have a PC with a fairly high storage capacity, I would like to set up an FTP server...

25 October 2016 9,769 5 View

Does TON (turnover number) value have any units?

Does TON in catalysis have any units? I have seen overlap of TOF and TON (TOF=TON/t), but papers seem to be using s-1 for both. I will be grateful if anybody can shed light on a proper...

07 October 2016 6,560 4 View

Is there a protocol for fluorescence titration for DNA-ligand interaction for determination of binding constants?

What is the difference between performing fluorimetric titrations for identification of binding stoichiometry and those for determination of binding constants? Is the titration of DNA to ligand...

17 April 2016 5,386 4 View

Can use D2O peak at 4.7 ppm to align 1H 1D NMR Spectra?

Hello all, I have taken the 1D proton NMR spectrum of my DNA sample in 90% H2O and 10 % D2O. I have also taken the spectrum of my sample with the ligand (dissolved in deuterated acetonitrile)...

13 April 2016 6,475 3 View

Is DNAse treatment of total RNA from MCF-7 cells absolutely necessary when using random hexamer kits for cDNA generation?

I am trying to generate cDNA from MCF-7 cells. So far, I have been able to extract total RNA using a combination of Trizol and Qiagen RNEasy kit. My A260/280 values are all above 2.0 and the...

30 July 2015 8,618 1 View

How much time does it take to perform a single UV-melting experiment to determine DNA Tm value?

I need to perform Tm for several sequences and need to provide an time estimate for it. My lab is considering procuring a new UV-spec so I would like to know how much time it takes per sample and...

18 June 2015 3,647 3 View

Is there a standard protocol for Tm analysis using CD melting data?

I am trying to find a protocol for performing CD melting experiments on oligonucleotides to analyze their Tm values. All papers mention that they collect the ellipticity at a specific wavelength...

23 February 2015 5,957 4 View

How can a difference in GC content in PCR primers affect PCR?

I have a fwd primer with 37% GC content and the rev primer with 61% GC content. The Tm of the fwd primer is 55.6 C and that of the rev is 58.2 C. Upon performing gradient PCR, I saw bands at 50C...

25 December 2014 3,764 8 View