Monika Gostic

10 Questions 12 Answers 0 Followers

Questions related from Monika Gostic

In situ hybridization protocol for SKIN cells (zebrafish)?

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 9,861 1 View

What would be the best marker to label ALL (or as many as possible) glia cells in zebrafish?

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 5,605 1 View

Fixing whole mount transgenic zebrafish for Imaging analysis?

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 8,894 4 View

Long range PCR optimisation?

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 7,531 8 View

Evaluating Tol2 F0 results in zebrafish?

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 5,790 8 View

What do I do when my RNA purity is over 3?

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 6,202 13 View

How do I obtain a high concentration of my final antisense probe for in situ hybridization?

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 1,882 2 View

How do I see how the change in amino acid changes the function of the protein?

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 4,009 13 View

What is the best positive control for in situ Hybridization of early developmental genes in Zebrafish

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 9,310 7 View

How do you insert a PCR fragment in reverse direction into the vector?

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 552 5 View