Nikki Schütte

5 Questions 4 Answers 0 Followers

Questions related from Nikki Schütte

How to make conditioned medium?

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 6,372 4 View

How can I obtain pure colonies from suspension (Jurkat) cells?

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 9,247 0 View

How to make limiting dilution with Jurkat suspension cells?

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 4,912 5 View

What ratio of gRNA to Cas9 do I need?

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 8,584 4 View

Is cleanup after ligation necessary, if I will be doing a successive ligation?

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 5,584 3 View