Sigrun S.

6 Questions 3 Answers 0 Followers

Questions related from Sigrun S.

Relation of EdU and FxCycle/7AAD/DAPI labelling?

Hi folks! I've been struggling a bit with understanding cell cycle assays, i.e. EdU assays. We do them in our lab with the click-it kit from thermo, using EdU and FxCycle (stains total dsDNA)...

08 August 2018 9,787 2 View

Alternative to sonication and benzonase for lysing DNA in WB samples?

Hi everyone, I'm looking for an alternative to sonication or benzonase for lysing DNA in Western Blot samples. Briefly, I am performing a subcellular fractionation on 3 Mio cells with a...

10 October 2017 7,579 2 View

T-cell Activation: Lck and p44 downregulation / degradation?

Hi guys, I am hoping to get some advice / insights into something I'm seeing in the lab. So I'm activating Jurkats with anti-CD3 and anti-CD28 antibodies for 0-10 minutes. After cell lysis I...

11 November 2016 9,351 2 View

Linearised plasmid obtained after Miniprep?

Hi everyone, We are trying to do a pretty simple restriction digestion but are having some issues and think it might be due to the plasmid we are trying to digest. Briefly, the plasmid is about...

07 July 2016 1,850 3 View

Can liquid E.coli cultures be frozen for plasmid purification?

Hi everyone, We have some liquid cultures growing that we would like to use for both making long-term glyercol stocks and for purifying plasmids from them. In between the liquid culture being...

06 June 2016 7,642 4 View

Using Restriction Enzymes in different companies' buffer?

Hei everyone, We are starting a new project and realized we have quite a lot of restriction enzymes left from a previous one. We are planning to do two double digestions and then ligate the...

06 June 2016 6,049 2 View