I have to Knock-out a gene using TALEN. So, its rather big plasmid.
Dear Huang, Thanks for the suggestion. I will see if it works.
I am using DMEM from gibco for the culturing, but cells seems not happy. I have also tried DMEM Glutamax but in all case cells do not survive more than one month. And the growth is very slow. Is...
07 August 2014 5,886 9 View
Could anybody suggest how to make MEFs transiently transfected by PEI method?
08 September 2013 9,533 1 View
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
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
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
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
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
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
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
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
My baseline increases without any explanation, it is only fixed by changing the solution B, will it be the column? Reagents? Bomb? Detector? Solution A: H3PO4 0.01% IN WATER Solution B: H3PO4...
21 February 2021 5,656 3 View
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
