Anil Kiran Chokkalla

19 Questions 5 Answers 0 Followers

Questions related from Anil Kiran Chokkalla

I need a tested RNA dot blot protocol!

I am doing RNA dot blot experiment to probe epitranscriptomic changes in brain tissue. I am not getting good signal with total RNA. I doubt my denaturation step as I am heating my samples at 95...

07 July 2017 9,476 1 View

Does reverse transcription of circular RNA yields a circular DNA with random hexamers?

I have seen many papers performing divergent PCR to detect the circular RNA products. I am not sure if the reverse transcription of circular RNA yields circular DNA with random hexamers.

06 June 2017 9,476 0 View

Why do I get a contamination in my H1 cells?

I am working on H1 human embryonic stem cells and culturing them on matrigel coated plates. On the second day of passage, I could see the media appearing cloudy and plate looks grainy with a lot...

07 July 2015 3,624 8 View

Any suggestion on a problem in sub-culturing H1 human embryonic stem cells with feeder free culture conditions?

I am working on H1 cells, I am sub-culturing in 10 cm dish coated with matrigel at 1:10 split ratio of confluent cells. I am having this frequent problem of cells not getting colonized on 3rd of...

06 June 2015 6,943 2 View

What is the best reference gene for normalising the expression level of a given gene of interest in qPCR experiment?

I was trying to analyse expression levels of certain genes after treatment of monocytic THP1 cell line with antigenic proteins through qPCR and i tried using the reference genes beta actin and...

08 August 2014 5,068 7 View

Does anyone have a composition of manually made TRIZOL reagent that worked well?

I am trying to isolate RNA from THP1 cell line using manually made TRIZOL reagent, I am getting good intact RNA but along with it I am getting huge DNA contamination. So I want to somehow minimise...

08 August 2014 5,095 8 View

Which is the most reliable method for quantification of gene expression data in qPCR experiment?

There are many methods for quantification of gene expression (fold change calculations) through real time PCR such as Pfaffl method, delta ct method, standard curve method. So I want to know which...

08 August 2014 2,173 4 View

How does the interaction between eRF3 and PABP stabilize mRNA during translation termination?

mRNA stabilization as a result of translation termination.

11 November 2013 7,771 2 View

How does the interaction of exon junction complex (EJC) and Upf proteins with translation termination factors lead to mRNA decay?

I want to know the reason why interactions between translation termination factors and EJCs lead to mRNA decay during NMD.

11 November 2013 6,153 0 View

What is the utility of real time PCR over northern blot?

Utility of real time pcr over northern blotting

11 November 2013 6,188 3 View

What is the purpose of circularization of mRNA in eukaryotes?

Seeking to learn the mRNA circularisation significance.

10 October 2013 4,858 1 View

Why do bacterial genes undergo Nonsense Mediated mRNA Decay (NMD) when introduced in eukaryotes even though NMD is not conserved in prokaryotes?

Nonsense mediated mRNA decay of bacterial genes in eukaryotes.

10 October 2013 3,917 3 View

What is the difference between IRES and kozak sequences?

Difference between Internal Ribosome Entry Sites and kozak sequences.

10 October 2013 759 4 View

How Exon Junction Complex (EJC) together with Upf proteins trigger non-sense mediated mRNA decay?

I want know exact mechanistic model of EJC, Upf proteins triggering NMD.

10 October 2013 6,728 2 View

What is the purpose of ATP dependant RNA helicase activity of Upf1 in non-sense mediated mRNA decay(NMD)?

The role of RNA helicase activity of Upf1 in NMD.

10 October 2013 2,879 4 View

How does Poly(A) at 3' end of mRNA confer stability to mRNA?

What is the role of Poly(A) in conferring mRNA stability?

10 October 2013 3,378 4 View

What is the difference between translational termination in normal mRNA and PTC bearing mRNA?

I want to know exact differences in remodeling events in case of normal translation termination and aberrant translation termination PTC bearing mRNA.

10 October 2013 5,763 0 View

Is MDA-MB-453 cell line triple negative?

I am studyingcombination therapies in TNBC and I want to know weather MDA-MB-453 belongs to TNBC sub type.

09 September 2013 3,162 13 View

Are the BT-20 cancer cell line Anchorage dependant?

To know whether colonies can be formed on soft agar plate.

09 September 2013 5,589 0 View