Mansi Srivastava

13 Questions 14 Answers 0 Followers

Questions related from Mansi Srivastava

Low library yield using Illumina True seq small RNA library prep ?

I am looking for suggestions to improve the yield of my libraries prepared using Illumina True seq small RNA library prep kit. Input RNA is purified small RNA (50 ng) and I run for 15 PCR cycles....

02 February 2020 8,681 2 View

How much time is required by RNase to optimally degrade RNA from a protein-RNA sample?

I am working on RNA-protein adducts, and trying to get rid of the unprotected RNA regions (that is not occupied by any protein). I am using RNAse A/T1 to degrade the RNA. For how long should I...

09 September 2019 3,794 2 View

How to increase RNA yield after ribosomal RNA depletion?

Hi all, I am working with small RNA fragments (30-50 bases). After ribosomal RNA depletion, I am exausting almost all of the RNA and there seem to be almost negligible yield of RNA (starting...

08 August 2019 8,638 3 View

Why my single stranded RNA bands don't appear on a denaturing agarose gel?

I am trying to check the size of my single stranded RNA on agarose gel with a denaturing loading dye in TAE buffer. Although ssRNA ladder bands are appearing but my samples (having 25-100 base RNA...

04 April 2019 310 3 View

Can anyone help me with troubleshooting my western blot with multiple bands in background?

I am trying to look at the expression of high molecular weight proteins NOS2 and p-NOS1 in raw macrophages but I see multiple bands in the blot with no clear bands of NOS2/p-NOS1 at their expected...

02 February 2016 1,245 12 View

Which is optimum cDNA concentration for RT PCR run?

Please suggest the optimum concentration of cDNA for RT PCR run. Does diluting the cDNA 1:10 or 1:100 help in increasing amplification, specially for low expressing target genes?

01 January 2016 8,731 5 View

Is Ct values above 35 in RT PCR run acceptable or not?

Can anyone suggest me whether Ct values above 35 in RT PCR run is acceptable or not? And whether it signifies primer dimer/ non-specific amplification. What is the acceptable range of CT values in...

01 January 2016 5,049 27 View

Why can I get no ct value for my RT PCR experiment?

I am trying to amplify cytokine genes in cDNA derived from macrophages but unable to get any amplification using comparative delta Ct experiment. Cross checked mRNA concentration and purity which...

01 January 2016 1,039 6 View

Suggestions on concentration of PMA to be used for differentiating THP1 monocytes into macrophages?

Kindly suggest me optimized PMA concentration that will work well for differentiating human monocytes THP1 into macrophages. I am facing problem with PMA concentration which is not helping THP1...

01 January 2016 1,855 3 View

Should the expression level of housekeeping gene vary with experimental condition or remain the same as that of control during RT PCR run?

I am using GAPDH as control housekeeping gene, to normalize the value of my target genes under experimental condition for real time run. I am getting variation in the Ct values of GAPDH in control...

08 August 2015 2,618 5 View

How can I increase the solubility of a compound for an MTT assay?

The compound used for MTT is not soluble in water and sparingly soluble in 5% DMSO, due to which there is issue with uptake in the cells. How can this be solved?

07 July 2015 7,686 15 View

How to remove protein contamination from RNA samples?

I have prepared RNA samples that seem to have protein contamination in them as the A260/280 ratio for them is between 1.5-1.7. Can I use proteinase in the sample and is there a way to get rid of...

07 July 2015 8,155 15 View

How can adherent cells be dual stained?

I am trying to dual stain live and dead cells (MCF7 and A375 cells). The problem is that after treatment of drug, the dead cells come into suspension and washed away after staining. Only few dead...

07 July 2015 1,754 6 View