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

How to design primers for miRNA quantification by qPCR?

If I want to design primers or mature form of a miRNA it is very short, so how to get primers. Can anybody explain in detail?

08 September 2013 809 2 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

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

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

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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

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

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

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

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28 February 2021 9,936 2 View

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View