Can anyone suggest transfection methods for big inserts in neuroblstoma or glioblastoma cell lines?

More Suparna Sengupta's questions See All

Can I use Sephadex G-200 to separate a complex of 2 Megadalton from a 100,000 protein?

I have 1 ml of 0.4 mg protein which I want to separate. Mainly I want the complex which should come in the void volume. I used a 40 ml bed volume manually packed column (0.75 cm diameter), but...

05 June 2018 4,767 3 View

Does anyone know why the retroviral shRNAs do not work at a later time point?

We are using retroviral shRNAs (both tagged and untagged) and made glycerol stocks after getting them from Origin. Have used them a number of times with proper downregulation profile. But now they...

03 April 2018 4,255 3 View

Can anybody suggest some adherent cancer cell lines with high intrinsic c-myc levels?

I want to study the role of c-myc in some downstream processes. For that I need to start with cells intrisincally overexpressing c-myc. Can anybody suggest some adherent lines?

11 December 2014 7,564 6 View

Is there any cell lysis buffer which will extract only cytosolic proteins, leaving other organelles?

I want to avoid centrosomal proteins.

10 November 2013 4,662 5 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

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

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

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

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

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

Why the cells die after stable transfection?

I tried stable transfection in the U87 cell line with the sgRNA-Cas9 All-in-One lentivector. Cells were cultured in a 6-well plate, 4ug plasmid was transfected after 24hs when the cell confluence...

22 February 2021 2,864 4 View

Is there a method for evaluating nanoparticle adhesion to epithelial tissue in vitro?

I want to measure how well nanoparticles can transfect cells based on NP adhesion properties. In the literature I find people do it in animal studies, and epithelial adhesion helps in transfection...

21 February 2021 7,791 3 View

Are exogenous molecules transferred between human cells during doubling ?

Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...

21 February 2021 6,633 3 View

Which transfection reagent is the best for plasmid transfection of MDA-MB-231 cell line, Lipofectamine based or other reagent suggestion?

I try to transfect MDA MB 231 cell line with a plasmid by PEI, unfortunately, the rate of transfection is low, and now I wanna find and order the best transfection reagent for this cell line and...

19 February 2021 828 6 View

Guillaume Freund

Hi Suparna,

I would suggest testing jetPRIME, a powerful and versatile plasmid DNA/siRNA delivery reagent. Many cell lines including U-251 MG and IMR-32 have already been succesfully transfected using jetPRIME.

If interested, feel free to request a sample!

Best regards

http://www.polyplus-transfection.com/sample-request/

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