14 Questions 14 Answers 0 Followers
Questions related from Monika Gostic
I am searching for a good reference image that would cover brain, eyes, otoliths and one for notochortd and spinal chord? Thank you
10 October 2017 9,428 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,...
06 June 2017 8,038 1 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...
12 December 2016 1,585 6 View
I am creating an in situ hybridization probe and am linearizing 10ug of plasmid DNA containing my construct. Would you clean up the product before using it for T7 transcription or would you use it...
10 October 2016 10,103 6 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...
08 August 2016 2,214 9 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...
08 August 2016 2,463 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...
04 April 2016 8,760 9 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...
03 March 2015 410 14 View
I have a final Gateway Tol2 construct ready for microinjection into zebrafish embryos. I also have a pCS-zT2TP zebrafish transposase mRNA, which I will co-inject with the construct. The question...
11 November 2014 2,373 2 View
I have an mCherry signal in my zebrafish and would like to detect it with antibody, which will enhance the signal. Does anyone have any recommendations? Also - can I use a mouse anti-mCherry for...
11 November 2014 7,599 7 View
I'm performing Tol2 enhancer and promoter assay in Zebrafish and would need a help with the constructs. I've ordered few of them from Addgene and they are on their way (for promoter assay: p5E-MCS...
09 September 2014 2,346 3 View
I am preparing a methanol stock of fixed embryos at different developmental stages (up to 5dpf) and storing them at -20C until further use. I fix embryos in 4% PFA in DEPC-PBS at 4C over night,...
09 September 2014 6,358 3 View
I'm performing the Gateway Tol2 transposition in zebrafish and am currently at the point where I would need to co-inject the destination vector with my construct and transposase mRNA. Does anyone...
09 September 2014 6,055 1 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...
08 August 2014 2,871 6 View
