Lech Kaczmarczyk

8 Questions 32 Answers 0 Followers

Questions related from Lech Kaczmarczyk

What are the possible ways to improve recombination efficacy of Cre- and Flp- dependent STOP cassetes?

Cre recombination efficiency often impacts activation of Cre/LoxP dependent transgenes. We have recently encountered problems with recombination in certain cell types in vivo, especially with...

12 August 2019 7,587 3 View

Can MeDIP-seq (methylated DNA immpunoprecipitation sequencing) be performed on DNA previously stained with DAPI or Hoechst?

Hi All, Can MeDIP-seq (methylated DNA immunoprecipitation-sequencing) be performed on DNA previously stained with DAPI or Hoechst? If not, are there any DNA stains compatible with the...

14 November 2016 2,658 1 View

What is the function of sucrose in lysis buffer for isolation of intact nuclei?

What is the function of sucrose in lysis buffers for isolation of intact nuclei? I observed that nuclei pellets resuspend easier in buffers without sucrose, and have a higher tendency to clump...

15 June 2016 8,586 10 View

I'm looking for a good antibody against mouse nuclei, suitable for FACS - any recommendations?

Hi All, I'm looking for an antibody directed against mouse nuclei (not cell type-specific), preferably directed to component(s) of nuclear envelope, and being a monoclonal. Antibody is intended...

07 June 2016 4,218 1 View

What is the best strategy for FACS staining of nuclear protein for sorting fixed nuclei?

Hi All, I need to FACS-sort formaldehyde-fixed nuclei from mouse brain tissue. What is the best strategy for staining of intranuclear protein for immunolabeling for FACS (staining of soluble...

15 May 2016 5,318 3 View

Is it possible to use Miltenyi MACS micro Beads and/or uMACS beads in tubes instead of columns?

Hi All, I was just wondering if anyone tried to use any of the Miltenyi beads in tubes instead of columns, with a standard magnets compatible with the "tube" magnetic separation format. My guess...

04 May 2016 330 3 View

Can Taq polymerase be used in SOE PCR?

Should using Taq polymerase be avoided when when doing SOE PCR? I would assume that, due to addition of non-template A and lack of exonuclease activity, it would generate mutant extension...

19 July 2015 4,173 5 View

What is the optimal concentration of SCR7 inhibitor for gene targeting in ES cells using CRISPR/Cas9?

Anyone has experience with using different concentrations? What are the potential adverse effects of (too) high concentrations?

12 June 2015 2,064 8 View