Subramaniyam Ravichandran

7 Questions 16 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 4,088 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 6,283 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 6,302 3 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 6,381 4 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 8,571 1 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 5,899 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 3,598 8 View