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...

06 July 2017 9,436 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.

05 June 2017 9,438 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...

06 July 2015 3,586 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...

05 June 2015 6,906 2 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...

07 August 2014 5,036 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...

07 August 2014 2,115 4 View

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

07 August 2014 5,032 7 View

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

mRNA stabilization as a result of translation termination.

10 November 2013 7,737 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.

10 November 2013 6,120 0 View

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

Utility of real time pcr over northern blotting

10 November 2013 6,143 3 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

To do liquid biopsy and look for specific gene expressions there are so many published papers. Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from...

23 February 2021 7,326 5 View

Universal bacterial primers targeting a mRNA?

Hi, I'm looking for universal primers targeting a bacterial mRNA to quatify bacterial load by qPCR in a complexe mixture (eukaryote and prokaryote RNA). Ribosomal RNA being depleted, I absolutly...

22 February 2021 9,969 3 View

How can I get the coding gene sequences of Hela and U2OS cell lines? Is mRNA sequence of Hsp90 alpha tissue specific??

I am looking for hsp90 mRNA sequence in Hela and U2OS cell lines, I have tried through ncbi but always the output is hsp90 in homo sapiens not in the specific cell line. Kindly, could any one...

22 February 2021 1,853 3 View

Can RNAscope be used to quantify the mRNA expression level of certain genes?

Hello, all, I want to explore the mRNA expression level of certain genes in the brain under physiological and pathological conditions. In previous studies, the researchers usually isolated the...

18 February 2021 1,518 1 View

In what ways do fellow reaerchers think that mRNA vaccination may be under CRISPR systems technology application ?

There are many ways for mRNA to intervene with genome expression. This question is raised for experts in CRISPR systems. Any RNA does not stand alone within acell. There are many ways to become...

18 February 2021 9,109 5 View

Wat is the half-life of luciferase protein in mouse tissue when delivered using mRNA luciferase?

Hello, I I am validating a system to deliver mRNA using nanoparticles to a mouse model. I am initially using mRNA for firefly luciferase to be able to do live imaging in the mouse. After that, I...

17 February 2021 2,536 3 View

Can I express protein of plant origin in E. coli strain (M15) using pET28a vector systems? Are M15 strains and pET vector systems compatible?

Are E.coli strains M15 and pET28a vector systems compatible? Can I over-express my protein which is of plant origin. Previously I was using BL21 DE3 strains. Was confused to use M15 strains for...

10 February 2021 2,564 3 View

Flexibility of RNAseq?

When you use RNAseq I understand that it must follow certain conditions when you use it. One of them is the insertion of a polyA tail. The insertion of the polyA must include then...

05 February 2021 3,770 4 View

Safety of the SARS-Cov-2 mRNA vaccine?

Dear colleagues I have the following questions concerning the safety of mRNA vaccines targeting the spike protein of SARS-Cov2: 1) The vaccine elicits antibodies which block the peptide domain...

04 February 2021 721 6 View