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Zebrafish Tol2 Gateway-compatible Kit for Neurodegenerative Disorders
(Kit # 1000000087 )

Depositing Lab:   Nicholas Cole

This kit is designed to streamline the production of transgenic zebrafish for studying neurodegenerative disorders. The kit contains 19 plasmids consisting of four promoters, six fluorophores, and empty vectors with MultiSite Gateway® (Invitrogen) compatible cloning sites or multiple cloning sites for the easy insertion of genes of interest.

This kit will be sent as bacterial glycerol stocks in 96-well plate format.

Add to Cart
$375 USD + shipping

Available to academics and nonprofits only.

Original Publication

A Tol2 Gateway-Compatible Toolbox for the Study of the Nervous System and Neurodegenerative Disease. Don EK, Formella I, Badrock AP, Hall TE, Morsch M, Hortle E, Hogan A, Chow S, Gwee SS, Stoddart JJ, Nicholson G, Chung R, Cole NJ. Zebrafish. 2016 Sep 15. [Epub ahead of print] doi: 10.1089/zeb.2016.1321 PubMed PMID: 27631880.

Description

In order to streamline the production of transgenic zebrafish for the study of neurobiology, the Cole Lab has designed a Tol2 Gateway compatible toolbox based on the original Tol2kit generated by the Chi-Bin Chien lab (Kwan et al., 2007). This collection, deposited by the Cole lab contains a set of 19 vectors.

This kit contains empty vectors with MultiSite Gateway® (Invitrogen) compatible cloning sites or multiple cloning sites for the easy insertion of genes of interest. Additionally, there are plasmids containing:

  • 4 promoters for differing expression:
    • motor neuron
    • pan neuronal
    • microglial/macrophage
    • astrocytic
  • 6 fluorophores
    • mTagBFP
    • TagRFPt
    • EGFP
    • mVenus
    • mCerulean3
    • mKOFP2

These vectors can be easily recombined using MultiSite Gateway® Three-Fragment vector construction protocols. The final constructs can then be used to create transgenic zebrafish for the study of neurobiology and neurodegenerative diseases.


Zebrafish Tol2 Toolkit Overview

An example of a transgenic zebrafish created with the Cole Lab toolbox to study the relationship between neurodegenerative disease and motor neuron degeneration. Two constructs are described here from the entry vector elements collated in this toolbox: pTol2pA2--3mnx1:mTagBFP, pTol2pA2--3mnx1:mCherry. Each expression cassette has a three prime SV40 polyadenylation signal (pA) in either the 3′ vector or backbone to aid expression.

How to Cite this Kit

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.

For your Materials and Methods section:

“The Zebrafish Tol2 Gateway-Compatible kit was a gift from Nicholas Cole (Addgene kit #1000000087).”

For your Reference section:

A Tol2 Gateway-Compatible Toolbox for the Study of the Nervous System and Neurodegenerative Disease. Don EK, Formella I, Badrock AP, Hall TE, Morsch M, Hortle E, Hogan A, Chow S, Gwee SS, Stoddart JJ, Nicholson G, Chung R, Cole NJ. Zebrafish. 2016 Sep 15. [Epub ahead of print] doi: 10.1089/zeb.2016.1321 PubMed PMID: 27631880.

Tol2 Gateway compatible toolbox for the study of the nervous system and neurodegenerative disease - #1000000087

Resistance Color Key

Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.

Inventory

Searchable and sortable table of all plasmids in kit. The Well column links to its kit plate map on the right. The Plasmid column links to a plasmid's individual web page.

Kit Plate Map

96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.

Resistance Color Key

Ampicillin
Kanamycin

Inventory

Well Plasmid Resistance
A / 1 pTol2p2A--3mnx1:mTagBFP-NS-MCS
Ampicillin
A / 2 pTol2p2A--3mnx1:loxP-mTagBFP2-stop-loxP-mCherry-NS-MCS
Ampicillin
A / 3 p5E-mpeg1.1
Kanamycin
A / 4 p5E-gfap
Kanamycin
A / 5 p5E-elavl3
Kanamycin
A / 6 pME-mTagBFP
Kanamycin
A / 7 pME-mTagBFP-CAAX
Kanamycin
A / 8 pME-mVenus-CAAX
Kanamycin
A / 9 pME-H2B-mCerulean3-P2A-mKOFP2-CAAX
Kanamycin
A / 10 pME-mKOFP2-CAAX
Kanamycin
A / 11 pME-H2B-mCerulean3
Kanamycin
A / 12 pME-loxP-mTagBFP2-stop-pA-loxP
Kanamycin
B / 1 p3E-MCS
Kanamycin
B / 2 p3E-mTagBFP
Kanamycin
B / 3 p3E-mTagBFP-CAAX
Kanamycin
B / 4 p3E-H2B-mCerulean3-P2A-mKOFP2-CAAX
Kanamycin
B / 5 p3E-mCherry-NS-MCS
Kanamycin
B / 6 p3E-EGFP-NS-MCS
Kanamycin
B / 7 p5E--3mnx1
Kanamycin
Data calculated @ 2017-09-25

Kit Plate Map - #1000000087

Plate #1

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