Is it possible to validate a miRNA target in a test tube by providing all the essential factors, and how can we determine the half life of a miRNA?

To validate a miRNA target mRNA can we do it in vitro by adding everything in a test tube? If not, what are the difficulties and how can we overcome this?

08 September 2014 925 4 View

Can anyone explain how to measure mitochondrial copy number using realtime PCR approach?

06 July 2014 7,576 8 View

Can anyone explain why my MTT assay is giving off a brown colour instead of purple?

When I perform an MTT assay after adding DMSO, only a brown colour is appearing. I am working with suspension cells and I want to know the cytotoxicity of a compound. After 24 hours of seeding...

11 December 2013 7,245 15 View

Can anybody explain how to design primers for a mature miRNA ?

I want to check the miRNA levels and miRNA transcript is short in length. How to design primer for a specific mature miRNA?

11 December 2013 9,870 7 View

Can anybody give me protocol for mitochondria staining using mitotracker?

I want to stain the mitochondria in suspension cells by using mitotracker red (Invitrogen). Can anybody explain protocol to observe them in fluorescence microscope?

11 December 2013 761 12 View

Why is protein aggregating during concentration by amicon?

Buffer composition is 50mM tris pH7.5(p.I 6) 150mM Nacl (tried upto 300mM) 5mM BME 1mM pmsf 10% glycerol. Even it is also aggregating after NI -NTA purification but I am diluting protein then it...

11 December 2013 9,397 6 View

Can anyone please recommend a protocol for analysis of surface expression of a CD marker using flow cytometry?

When I am performing an experiment I could not observe the any signal. Usually I use FITC tagged antibody but I am suspecting that I have problem with sample preparation and cells may be damaging.

08 September 2013 8,043 14 View

Does anyone have CMK cell line?

I am working on the megakaryocyte development. I need this cell line for my study.

02 March 2013 5,278 1 View

Can anyone send me a detailed procedure of miRNA cloning in a mammalian expression vector?

Can anyone send me a detailed procedure of miRNA cloning in a mammalian expression vector?

11 December 2012 594 0 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

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 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

Can MicroRNA cross the blood brain barrier?

In diseased conditions, many genes and gene products are visible in the bloodstream. So my question is whether we can find traces of microRNA which is involved in disease and disorder in blood serum?

24 February 2021 7,064 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

Are exogenous molecules transferred between human cells during doubling ?

Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...

21 February 2021 6,633 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