Is there a good reference image for 0-5 dpf zebrafish?

I am searching for a good reference image that would cover brain, eyes, otoliths and one for notochortd and spinal chord? Thank you

09 October 2017 9,302 2 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,...

05 June 2017 7,937 1 View

Zebrafish mounting for confocal microscopy?

11 December 2016 1,494 6 View

Would you usually clean up linearized plasmid before using it for ISH probe?

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...

09 October 2016 9,976 6 View

Long range PCR optimisation?

07 August 2016 2,098 9 View

Up-to-date CRISPR knock-out protocol in zebrafish?

07 August 2016 2,347 6 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...

03 April 2016 8,648 9 View

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

02 March 2015 308 14 View

Can anyone suggest a good anti-mCherry antibody?

10 November 2014 7,500 7 View

Any advise on the Gateway Tol2 Constructs?

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...

08 September 2014 2,205 3 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Anyone know of a good progesterone antibody?

I am looking for a good progesterone antibody to quantify levels of progesterone in zebrafish. Applications would be ELISA, and immunohistochemistry.

28 February 2021 5,929 2 View

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View