Sigrun S.

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Questions related from Sigrun S.

Two sgRNAs 500bp apart --> deletion?

Hi everyone, I am trying to knock out a non-coding region downstream of my gene of interest. I looked at a publication which did exactly this but in a different species. They seemed to use two...

25 June 2022 9,587 3 View

Probe-based DNA enrichment - how to avoid re-hybridization with complementary DNA strand?

Hi, Quick conceptual question; in any assay that relies on enriching DNA with an oligo probe, e.g. having a biotinylated oligo that can be pulled down with streptavidin coated beads; How would one...

16 July 2020 6,823 0 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...

18 October 2017 9,554 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...

07 November 2016 6,330 2 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...

22 June 2016 9,380 4 View