17 Questions 8 Answers 0 Followers
Questions related from Beatriz María Fernández Santos
Hi, everyone Has anyone tried both Citrate Buffer pH 6 and TRIS-EDTA buffer pH 9 for inmunohistochemistry using autoclave and seen differences in the DAPI staining? It seems to get blurrier /...
4 weeks ago 8,899 3 View
Dear everyone, We've been dealing with EndoCBH1 cells for a while now. And even using their specific recommended cell culture media from Human Cell Design (OptiBMax) they are very hard to...
25 October 2024 8,868 1 View
Hi!! We've been using this product for quite a while now for our whole embryo in situs hybridization with mice embryos: https://www.sigmaaldrich.com/ES/es/product/roche/11383213001 The thing...
31 May 2022 3,870 0 View
Hi there!! If I have a colony with a plasmid that I've transformed onto it... is it possible to modify my plasmid with CRISPR? So that I can now purify a new modified plasmid? Is this...
23 September 2021 2,182 1 View
Hi everyone We've been trying to sequence our region of interest from some patients. The genomic DNA they've sent us is not of great quality, but PCR works so they musn't be terrible. I amplify...
30 August 2021 3,463 2 View
Hello everyone I have never cultured cells in my life and everything is new to me. I've done some tests with Lipofectamine® 2000 in my IMR-32 cell line with some plasmids containing dTomato that...
20 August 2021 7,418 7 View
Hi everyone We need Fast Green to electroporate plasmid into mice embryos, therefore we simply need it to see some color in the liquid we are electroporating so that electroporation is more...
08 February 2021 2,192 2 View
Hi there I'm planning on annealing two PCR products to perform some mutagenesis in a plasmid. I was recommended to try and keep it to 600pb each, for example, but for one of the plasmids there...
16 November 2020 9,012 3 View
Hi, there We are having some trouble trying to combine immuno-based colorimetric assay and methylene green staining, since we perform the inmuno as usual and then when we try to get methylene...
11 November 2020 5,248 1 View
Hello everyone We've been struggling with our project due to low efficiency rates + check the insert is OK and not in reverse + PolTaq did not introduce a new mutation... and it has come to our...
24 January 2020 6,238 3 View
Hi, everyone I am using a 4.3kb vector to clone both a promotor and an enhancer. I already have a positive miniprep with no mutations in the promotor, therefore I took some of this plasmid,...
09 August 2019 2,806 3 View
Hello everyone I've been trying to clone a 1640 pb insert into a 4300pb vector. Transformation works great, after O/N incubation at 37ºC, lots of colonies are found both in the control and in the...
20 July 2019 6,318 7 View
Hello everyone, We are trying to get some constructs after ligation of the plasmid and the insert of interest, and our efficiency rate is very low. Out of 10 colonies, usually only 1 or 2 (or...
16 July 2019 4,069 4 View
Hello there, Has anyone transfected anf of these cell lines and can suggest a protocol? We need a cell line that expresses a certain gene to try our fluorescent construct on, so we don´t mind...
22 March 2019 4,660 1 View
Hello there When we want embryos for cryostat, we usually wash them 3 times in PBS to remove PFA 4% and then we do sucrose 7.5% for a while at room temperature (basically until they sink) +...
22 February 2019 4,786 3 View
Good morning, everyone Yesterday I was performing a manual miniprep. One of the steps consists of a 4ºC max speed (in my case 18.000 rpm) centrifugation for 10 min. When it was done, all of the...
20 February 2019 8,165 1 View
Hello, everyone I've been struggling for almost a month already when amplifying a promoter from the whole mouse genome: - I already designed 4 primers and have been trying them all (meaning a...
29 November 2018 410 3 View
