Transfected cells died after trypsinization ?

Hi all, I am facing a problem to trypsinize and combine/move the transfected cells into new well . I was planning to pool the transfected cells into a new well to ensure better cell-cell...

11 December 2019 3,243 5 View

Constitutive expression of eGFP in transfected cells?

Hi, I am trying to culture a stable clones using transfected cells. As the clones grow, I noticed the eGFP signal diminishes over time but the cells are still resistant to antibiotics and do not...

11 December 2019 176 14 View

New in lentivirus transduction work?

Hi all, I am new in lentivirus transduction work and have some problem as listed below: 1) Is CMV promoter considered a strong promoter to express the gene I want in the transgenic organism...

09 October 2019 8,592 4 View

Problem with stable cell line generation?

Hi all, I am facing a problem to generate a stable cell line after successful transfection. I added puromycin to kill the untransfected cells, however, eventually all my transfected cells are...

08 September 2019 9,024 2 View

Will miRNA transfection affect cell morphology?

Hi Scientist, I am transfecting an oncogenic miRNA into a breast cancer cell line and based on your experience in the mammalian cell line, will such transfection affect the cell morphology?

06 July 2019 7,378 8 View

Suggestion for good plasmid extraction kit?

Hi, Can anyone recommend a plasmid extraction kit that could produce good plasmid yield (>300ng/uL). The plasmid I am using now is a high copy number plasmid and the bacteria I am using is...

06 July 2019 6,820 3 View

MiRNA primer sequence?

Dear all, I have a miRNA primer sequence left by someone else and I would like to know about the sequence of this primer. Is there any method I can use to sequence and know the sequence of this...

05 June 2019 5,879 7 View

Co-transfection of miRNA mimics and inhibitors?

Hi, Is there any paper which reported the co-transfection of miRNA mimics and inhibitors? Thank you.

05 June 2019 9,712 3 View

Use of fluorescence or luciferase approach in checking transfection efficiency?

Hi all, I would like to seek for your opinions on whether fluorescence approach (GFP) or luciferase approach is better in assessing transfection efficiency / success (after miRNA...

03 April 2019 1,383 6 View

Single plasmid that carries both miRNA and reporter gene like GFP/luciferase?

Hi researcher, I wonder any of you have tried to design a single plasmid that carries both miRNA and reporter gene like GFP / luciferase? I am thinking with such plasmid, I will just need to...

03 April 2019 5,144 7 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

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