What is wrong with the expression from pLKO lentivirus?

We obtained pLKO5-EFS-GFP from addgene (Ben Ebert Lab) which contains U6-gRNA-EF1s-GFP. Using transient transfection, we got very good transfection. With virus, the GFP expression is extremely...

06 July 2016 9,261 4 View

Is there any transgenic inducible CAS9 rat available?

I did not find any resource.

05 June 2016 7,552 3 View

How long does synthesized oligos last in cells?

if synthesized oligos are transfected into cells (in vivo), how long will they be stable? Are they going to be degraded within hours, days or weeks? Any information/references will be highly...

05 June 2016 6,690 0 View

Anybody used jetpei in vivo transfection in brain?

05 June 2016 6,775 2 View

Is EF promoter working well in neurons?

I am trying to subcloning the short EF1a promoter to drive Cas9 in neurons but not sure if it works as well as CAG promoter. Anybody has experience with EF1a in neurons?

02 March 2016 4,417 5 View

CAG promoter and CMV promoter with CAG enhancer are the same?

I am planning to express a gene(dCAS9) in neurons with CAG promoter based vector but only have a vector with CMV promoter with CAG enhancer. Are they the same regarding the expression efficiency...

31 December 2015 982 4 View

What's the best way to measure growth rates in House sparrow chicks from day 2 to day 10?

What's the best way to measure growth rates in House sparrow chicks from day 2 to day 10? Since, the growth curve from day 2 to 10 won't be like the "Logistic curve" it might not follow logistic...

03 March 2021 1,401 3 View

No title

02 March 2021 3,060 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

No title

28 February 2021 9,936 2 View

Can Ethovision store the generated data and video recordings directly to NAS system?

The size of data generated by Ethovision is huge. Having NAS system for data storage seems to be a good option, I am wondering whether Ethovision can directly store the data to the NAS system...

27 February 2021 6,350 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

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

What is the best surface to culture cells isolated from a skin biopsy?

Hello, recently I am having some difficulties with maintaining the cell culture of primary cells isolated from a skin biopsy (normal and malignant). The coating of culture plates with Matrigel (a...

26 February 2021 6,151 2 View

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View