Any idea how to troubleshoot my problem with target stable cell lines.?

Good day!I am currently generating stable cell lines( KON ) using TTA vector and G418 as drug selection agent.?After two weeks i got isolated colony from some wells at the 900ug/ml (G418). I...

02 March 2018 4,955 5 View

What is the best strategy to knock in cell line using cas9 system?

Is homology sequence recombination will work? Any construct suggestion. Thanks in advance..

02 March 2018 1,908 1 View

Good day! I am doing drug screening for Oral squamous Carcinoma cell Lines. My drug G418 concentration is from 100 ug to 100ug per ml?

It turns out that my cells become 100 percent (from 70% confluent) confluent the after 4 days? Any suggestion On optimizing drug screening.?

01 February 2018 4,753 2 View

Any idea regarding mutagenesis. Overlap extension PCR?

I am trying to amply a 1400bp gene.. My expected fragments which are 900 and 500 were achieved. I joined the two fragments and i got 1400 as expected. This is my mutation desire CAG to GAG....

31 December 2017 3,051 6 View

Good day. Anyone can help me regarding how much volume on the fragments should I use for overlap extension PCR?

The two fragments have the same intensity. Any detailed PCR Reaction for this. Thank you.

11 December 2017 6,925 3 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

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

No title

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