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Questions related from Monika Gostic
I have extensive experience in WISH and FISH in zebrafish embryos when it comes to mapping the expression of genes within the fish. I am now about to start a new project, checking the expression...
04 March 2020 9,943 1 View
I have conducted a study using Tol2 Gateway system to study human promoter in zebrafish. As per literature, the system should have worked, but it didn't. The zebrafish positive control was...
02 March 2018 2,949 2 View
I am identifying cells that express my gene of interest and I think they might be glia cells. I would therefore need a reliable marker that would label as many glia as possible - Muller,...
26 June 2017 5,646 1 View
I am doing an in situ hybridization and have localised my expression to the developing zebrafish eye. I would like to have some kind of positive control, but am not too familiar with eye...
07 June 2017 276 5 View
I'm doing a large scale analysis of expression patterns in transgenic zebrafish and I am chasing after certain developmental stages. Therefore I need to fix the fish at that stage and once the...
01 March 2017 8,950 4 View
I am performing a whole mount in situ hybridization on zebrafish embryos. I have increased hybridization T to 65C but am still getting nonspecific binding. Can I increase the T even higher and...
18 January 2017 7,658 9 View
I am trying to image my zebrafish embryos from 0-120hpf and am struggling with suitable mounting without damaging the specimen. Does anyone have a good protocol or own experience for it? I am...
01 December 2016 3,724 4 View
I'm using Tol2 Gateway system to test for human promoter activity in zebrafish. I am already using zebrafish UBI promoter as a positive control, but I was wondering if anyone might have a HUMAN...
24 October 2016 1,992 2 View
We're performing a long range PCR, longer than 3kb and so far it has failed to work. We've tried DMSO, Mg2+, touchdown approach, different DNA, new primers, different polymerases. I'd appreciate...
03 August 2016 7,583 8 View
Is anyone working with CRISPR/Cas9 in zebrafish and has a detailed up-to-date protocol with high efficiency? I would like to create a knock out of my protein of interest, but so far none of my...
03 August 2016 9,201 6 View
I am performing transgenesis in zebafish, using a Tol2 system. I have successfully constructed the entry plasmid and the integration of my wished construct has also been confirmed. However, I do...
29 April 2016 5,862 8 View
I'd like to visualize zebrafish cilia, for which I'd need an antibody such as the one against γ-tubulin. Does anyone know any good ones? Or if you might have other suggestions how to stain for...
12 June 2015 5,219 2 View
My CAP and PolyA RNA has been quantified on the nanodrop and all the samples show high concentration as well as the ration between 260/280. A is over 3 in all of them. Does that mean I have extra...
27 March 2015 6,275 13 View
I'm performing an In situ Hybridization on zebrafish and would need a probe for zebrafish otx5, expressed in pineal and parapineal. Does anyone have it (or maybe something similar). Many thanks!
05 March 2015 5,766 2 View
I've created a construct for Tol2 Gateway enhancer assay and am about to proceed with experiments. However, I do not have a positive control (nor negative). Can you suggest me what would be the...
26 January 2015 7,174 6 View
I created sense and antisense probe by extracting and purifying RNA, generating cDNA, performing PCR for wished construct, which I then cloned into pCR-Blunt II - TOPO vector. After isolating and...
08 January 2015 1,940 2 View
I have created capped, polyadenylated RNA in order to microinject it into zebrafish embryos. I would now like to make sure that what I've got is the right construct. How do I quantify the...
12 December 2014 9,456 2 View
I have a sequence of my gene and protein of interest and I have a database of mutants telling me where the mutation in the sequence is. I want to know how this mutation affects the protein...
07 November 2014 4,062 13 View
I'm performing an in situ Hybridization protocol and have no knowledge of the expression of my candidate gene. Firstly, I will perform a qPCR in order to see when the gene of interest is...
02 October 2014 9,366 7 View
I have a PCR fragment with blunt edges that I'd like to insert into my vector. I already inserted it in a normal forward direction, but now I'd like to try it also in the reverse direction. Does...
25 August 2014 601 5 View
I've taken over a project from a previous postdoc and have discovered a mutation on one of the sequences. This mutation is located right after ATG site and it mutates GCC into TCC, causing...
01 January 1970 9,928 0 View