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Questions related from Amila Zuko
I've uneven bands (size, intensity, and staining) when doing Western Blotting and I'm wondering what the problem could be. The protocol: -10% separating gel (0.375M Tris pH8.8; 0.1% SDS; 10%...
15 July 2019 6,753 8 View
I've performed microinjections to create a tag-insertion inside my gene of interest by CRISPR/Cas9. A quick PCR shows that a bunch of the pups are positive. However, all are heterozygotes. I'm...
23 February 2017 6,459 2 View
I'm performing IMAC to purify my his-tagged protein (enzyme) from e. coli lysate. It seems I can get a good amount of my protein of interest but the problem is that I also get alot of coeluates,...
13 October 2016 9,911 7 View
I have tried 6xhistagged-protein production and purification from E.coli but nanodrop couldnt detect any protein in eluates from nickel-resin beads. I suspect my problem is sonication: I spin...
29 July 2016 1,756 16 View
Several papers base their method of ES cell differentiation into neurons on Bibel et al., 2007 (Nat Prot). When the cells are initially plated for neuronal differentiation the medium contains N2...
23 June 2016 6,732 4 View
This concerns linear PEI 25kDa. So far I am aware of two methods: 1: stir the PEI in MQ (1mg/ml) and dissolve by dropping and holding the pH at 2. After its transparant increase pH to 7 -->...
21 April 2016 3,008 22 View
Lately I'm getting failed DNA sequences and strange sequence peaks in my chromatograms when trying to sequence my plasmid (see attachments). With a previous mini prep sample, the following...
09 February 2016 6,541 4 View
I would like to study the development of single cells (neurons) that were treated in vitro before transplantation into the mouse cortex. At the moment I have mouse embryonic stem cells available...
19 November 2015 9,717 4 View
I have been doing co-immunoprecipitations for a while now using my own buffers (CHAPS lysis buffer + protease & phosphatase inhibitors) and Dynabeads protein G beads. Useful information: -The...
22 January 2014 5,313 3 View