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Questions related from Nikki Schütte
I need to enrich for mitotic cells for immunofluorescence. Since the mitotic cells are rounded I need to increase the adherence to the surface of the plate. So far I have been using only...
29 November 2023 9,583 0 View
I need to do immunofluorescence on mitotic cells (HeLa). I coat the wells in poly-L-lysine diluted in H2O; washed 4 times with H2O. Cells are treated such that they arrest in the mitotic phase. A...
22 November 2023 5,564 0 View
I have run multiple agarose gels in which I have at least two combs. I continually see the lower part of the gel visualising much fainter than the top portion. The last band of my ladder (250 bp)...
15 September 2023 4,240 4 View
I need to prepare conditioned medium for Jurkat cells. I understand that it is medium in which the cells have already grown, and the cells filtered out, but how exactly should it be made? What...
01 August 2023 6,376 4 View
I am busy generating a KO cell line with Jurkat suspension cells. After transfection I perform single cell dilution (limiting dilution) to approx. 70-100 cells per 96-well plate. It seems that...
12 July 2023 9,252 0 View
12 July 2023 4,917 5 View
I will be generating a knock-in cell line using the following in transfection: Cas9:gRNA RNPs + 2.7kb dsDNA linear donor Attached is the protocol with the quantities required for each reagent...
07 June 2023 304 0 View
I am planning to generate a KO cell line, using RNPs I am using a paired gRNA system to induce a deletion of a segment instead of just indels. Generally the RNPs are formed using a 1:1 molar ratio...
10 May 2023 8,587 4 View
I have two linear DNA fragments that will be ligated together (each ~700 bp). This fragment then needs to be ligated into a linearised vector (plasmid). Would I need to perform a clean up on the...
08 May 2023 5,589 3 View
