Goyani Shanikumar

4 Questions 4 Answers 0 Followers

Questions related from Goyani Shanikumar

Which gene should I use as mitochondrial reference gene while doing qRT-PCR from mitochondrial fraction?

I am doing qRT-PCR to check the expression of certain miRNA and mRNA in the mitochondrial fraction. But I am not able to determine which reference gene I should be taking in the case of the...

20 June 2023 4,089 2 View

How can I calculate % input to plot RT-PCR data as graph in RNA-Immunoprecipitaton (RNA-IP) experiments?

I have recently done RNA-IP assay by pull-down of my protein of interest and my further applications involve the analysis of bound RNA to it. For this, I did RNA-IP and isolated RNA from the...

11 May 2023 344 0 View

Why does RT-PCR (Real-time) primers starts showing amplification in NTC (Non-Template Control) after using only a few times?

I am getting amplification and specific melt-curves in Non-Template Control sample (with water instead of cDNA). I ordered some primers and reconstituted them to 100 uM and diluted them to 10 uM...

08 April 2023 1,961 2 View

How can I clone shRNA for gene knock-down in pLKO.1 vector?

I have been trying to clone an shRNA for a gene knock-down in pLKO.1 vector but I am not able to get the clone. I don't know what the problem is. I have tried annealing the oligos at 95 C in a PCR...

30 September 2022 8,323 5 View