Normalization luciferase data assay with number of cells?

Hello everyone, I am running luciferase assay with pMIR REPORT plasmid to evaluate the function of a miRNA on a specific 3'UTR. I wondering if I could normalize the firefly signal to the number of...

06 July 2017 2,190 3 View

Wrong p53 size on Western blot?

Hi I am blotting for p53 in my experiment. The predicted MW is 53 kDa. However the actual MW appears to be higher than the expected MW in SDS-PAGE. I loaded 10 micro-gram protein. I didn't have...

06 July 2017 7,401 5 View

Identify cysteine residue that is responsible for disulfide bond formation?

04 May 2016 7,500 4 View

Can anyone tell me how to identify targeted proteins pertaining to apoptosis pathway?

Hi everyone, I am doing a project to predict and prioritize miRNAs and their targets that are involved in regulating apoptosis. We have already had the results of miRNAs expression and their...

01 February 2016 5,148 3 View

Antibody against pro-caspase 3 also recognizes same position of beta-actin?

01 February 2016 4,319 3 View

Is anyone familiar with postranscriptional regulation?

09 October 2014 4,425 10 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

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 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

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

What is the significance of (isolated) high titers of IgG anti GD1a antibodies in an older patient with polyneuropathy?

Remote history of Campylobacter infection with reactive arthritis, and Spitz nevus (UMP). Arthropod exposure in Zika/ChikV endemic area prior to emergence of length dependent, sensory PN with...

18 February 2021 8,719 1 View

Has anyone encountered any problems with abm good microRNA primers for quantification of mature miRNAs?

I've been trying to quantitate mature microRNAs in 293T cells with and without 'X'. 2ug of total RNA (isolated by Trizol method) is converted to cDNA using All-in one microRNA cDNA synthesis kit...

17 February 2021 8,986 2 View

It is easier to knockdown a gene than overexpress a gene when generate a stable cell line?

I am just so confused that some people tell me knockdown is easier than overexpression when generating a stable cell line. I think it depends on the gene. Cells won't be happy whether we try to...

15 February 2021 1,370 2 View

Would using an asRNA for repressing transcription of a high expression Listeria monocytogenes non-coding RNA harbored in a plasmid be a viable option?

Hello all, I am interested in deleting a highly expressed non-coding RNA located on a Listeria monocytogenes plasmid. Currently, there are not any optimized gene deletion methods for Listeria...

09 February 2021 4,994 1 View

What is the best way to fix plasma derived miRNA for subsequent cDNA synthesis?

I need to quantify blood plasma derived miRNA through q-RT-PCR. For this I need to fix RNA content in my cDNA reaction. What is the best way to do this? Currently, I take readings of my plasma...

03 February 2021 9,330 3 View

What is the general miRNA yield one gets from the Qiagen miRNEASY serum/plasma advanced kit?

I am using the Qiagen miRNEASY serum/plasma advanced kit, using 400ul of frozen plasma. What is the yield you get on average? I am using nanodrop currently for quantification. I am getting...

02 February 2021 8,806 2 View