Florian Pruckner

6 Questions 1 Answers 0 Followers

Questions related from Florian Pruckner

How to lyse nuclei without detergent?

Hi, I want to isolate nuclear proteins from the diatom P. tricornutum. I can successfully isolate the nuclei, and can image them under the microscope. But I fail to lyse the nuclei. As they were...

15 December 2023 3,675 1 View

What kind of linker to fuse mEOS4 to a protein of interest?

Hi all, I want to clone several constructs with a protein of interest fused with a mEOS4 protein. This will include N- and C-terminal fusion. Are there any linkers that are preferably used over...

28 April 2023 2,524 0 View

Which mEOS version to use for PALM?

Hi, I would like to use mEOS to tag proteins to evaluate their subcellular localization. I am new to PALM, and am a bit confused with all the different versions of mEOS available... mEOS2? mEOS4?...

20 April 2023 7,502 1 View

STORM microscopy to image sfGFP or mVenus?

Hi, I was wondering if it is known if STORM microscopy could be used to obtain super-resolution images of sfGFP or mVenus tagged proteins in cells. I know it is more common to use fluorescent...

17 February 2023 1,017 2 View

Does Filipin (fluorescent probe for sterols) staining kill the stained cell?

I want to stain a microorganism with a sterol specific fluorescent-dye for FACS and reculture high sterol producers after sorting. I know the fluorescent probe Filipin is available for staining...

10 January 2023 4,565 1 View

Are there known problems with golden gate cloning efficiency of large inserts (4300 kb)? How could GGC efficiency be improved?

Hi, I am having troubles cloning a Cas9 gene (around 4300 kb) into a vector, replacing a LacZ cassette. (I use the uloop system.) I use SapI as a restriction enzyme and T4 as a ligase. I cloned...

24 November 2022 6,606 1 View