What happens to the miRNA-RISC complex after it cleaves on a mRNA molecule?

I'm more focused on the miRNA than RISC. Does the miRNA degrade after cleaving mRNA?

09 October 2019 9,280 1 View

When the miRNA is bound to a mRNA, is it expressed in a miRNA PCR?

I conducted a miRNA PCR and I was wondering if miRNA that is bound to a mRNA is expressed in the results from my PCR. Thank you!

08 September 2019 7,420 7 View

Can pseudogenes affect miRNA gene expression or do they just competitively bind to miRNA?

The pseuodgene I'm researching has affected the miRNA levels in my cell culture (I found out through qPCR). However, I thought that pseudogenes just bind to the miRNA rather than affect its gene...

07 August 2019 1,959 2 View

What is the recommended concentration range of miRNA cDNA to use in a qPCR?

06 July 2019 4,290 5 View

How do you locate the exact spot of the scratch on a scratch assay?

I was wondering if anyone had any tricks to locate the exact spot of the scratch for a scratch assay. For example, I know some people in my lab have drawn a grid on their plates. Thank you!

06 July 2019 1,406 2 View

Will the PTENP1 plasmid transform properly in NEB 5-alpha Competent E. coli?

Addgene recommends the growth strain to be DH5alpha but we have NEB 5-alpha in my lab. Please let me know if my bacterial transformation can still work! https://www.addgene.org/35008/

11 December 2018 3,982 1 View

How do I get transfection efficiency with a plasmid with an HA tag?

I want to use a plasmid that is HA tagged. My research is in vitro and I am bit confused on what to do. Thank you!

08 September 2018 6,804 4 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

No title

02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 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

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View