Transcriptional or post-translational regulation?

I have came across a paper where they observed a delay timing of a drug inhibitory effect and insulin stimulatory effect respectively on purine synthesis, and thus, they suspected that the...

09 October 2018 9,919 0 View

How to process RNA-seq results?

I have obtained the RNA-seq results, which is in an excel file with all the fpkm values. May I know how do I check the gene expression profiles? Is there any software for this or I gotta manually...

04 May 2018 3,338 9 View

How to determine a protein is working downstream of another protein?

If overexpression of protein A can rescue the growth defect caused by mutation in protein B, can I consider protein A is working downstream of protein B? If yes, why is it so?

04 May 2017 7,522 24 View

Can anyone suggest method to extract membrane proteins?

Currently I am having problems in extracting membrane proteins (ammonium transporters), can anyone help? I used LDS in extraction and I did not heat the samples. Proteins in lane 1-4 seem like...

03 April 2017 773 20 View

Best temperature to dry DNA samples after ethanol precipitation?

I am preparing samples for BigDye sequencing. After PCR, I did ethanol precipitation to clean up the samples. In order to dry up the samples (after removing 70% ethanol), is it okay for me to dry...

09 October 2016 6,100 8 View

Overexpression of a gene is toxic to the cell? Why is it so?

Currently I am working on a new protein in fission yeast. Based on the predicted function, it is involved in transcription. We found that when this gene is overexpressed, the cell growth is...

06 July 2016 8,718 1 View

If one of the proteins in S.pombe was tagged with FLAG, is this pombe still considered as wild type?

One of the proteins in S.pombe that I am working on is tagged with FLAG, so can I still call this strain as wild type as it contains a fragment of foreign DNA? If no, what is the right term for it?

05 June 2016 8,096 3 View

Can I re-digest my digested PCR product?

I have digested my PCR product with NEB Not1 and Nde1 for 1hr. I have cut the band and purified the product. However I afraid the digestion is incomplete, can I re-digest them again? Is there any...

04 May 2016 2,655 17 View

Anyone has protocol for TaKaRa PrimeSTAR mutagenesis basal kit in English?

02 March 2016 4,844 6 View

What are the characteristics of a broad host range plasmid?

May I know what are the characteristics of a BHR plasmid? Is this kind of plasmid occurs naturally or we can try to engineer a narrow host range plasmid to become one?

01 February 2016 7,160 0 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

What is the exact mechanism of the antimicrobial effect of local anesthetics?

The antimicrobial effect of local anesthetics has been known for more than 50 years, but it still seems to have an unclear mechanism of action. which bacteria? which concentration? etc etc. ....

27 February 2021 4,450 2 View

When is it unnecessary to change organisational culture?

As a former management practitioner, I recall a generally accepted view that changing organisational culture is difficult, so you should only attempt it if the organisational change that you are...

27 February 2021 2,595 8 View