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

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

CGA 300 Regulator for CO2 cylinder?

Has anyone ever tried to deliver CO2 with a CGA 300 regulator? I know that a CGA 320 regulator is standard for CO2. The issue is that I currently do not have any free CGA 320 regulators. I also...

25 February 2021 3,292 3 View

RNA-seq DEGs downstream analysis and reporting in literature?

I used EdgeR to get the list of DEGs (differentially expressed genes) from my liver RNA-seq experiment. I have done the gene overrepresentation and GSEA based pathway analysis. I am currently...

23 February 2021 8,969 1 View

Could you please suggests a tutorial or paper or any other materials to learn multiple gene expression in TCGA and GTEx databases?

I would like to analyze multiple gene expressions in normal and cancer tissue by using the data in the TCGA and GTEx databases. Please suggest a tutorial or paper showing step by step analysis of...

23 February 2021 9,007 3 View

Are there any databases for gene co-expression or expression pattern clustering?

I am currently working on gene clustering based on co-expression pattern in mouse brain. The problem is I do not have some solid way to test my result. Are there any suggestions for databases...

22 February 2021 4,374 3 View

Which CpGs have a significant effect on gene expression?

It is known that methylated CpG around TSS decreases gene expression. However, It is difficult to know exactly which CpG has a greater impact on gene expression. Are there any databases known...

20 February 2021 5,634 3 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View

Purification of proteins from baculovirus-infected insect cells?

18 February 2021 6,665 6 View