[Zbrafish] Construct/transposase [ ]

Chi-Bin Chien via zbrafish%40net.bio.net (by chi-bin.chien from neuro.utah.edu)
Tue Jan 27 13:06:34 EST 2009


Hi Jonathan,

Those concentrations are reasonable, assuming that you are injecting  
about 1 nl/embryo. If you read any of the original papers on the Tol2  
transposon you will find details. For instance, look at our Tol2kit  
paper:

KM Kwan et al. (2007) The Tol2kit: a multisite Gateway-based  
construction kit for Tol2 transposon transgenesis constructs.  
Developmental Dynamics, 236:3088-99.

Or, read the Tol2kit wiki:

http://chien.neuro.utah.edu/Tol2kitwiki/

--Chi-Bin Chien

On Jan 27, 2009, at 7:56 AM, Jonathan Schumacher wrote:

> Greetings,
>
> I am a rookie with zebrafish transgenics and have been given a project
> to inject embryos with a construct in conjunction with transposase
> cRNA.  I don't see in the literature where the concentration of
> construct & transposase are described.  I've been using 20ng/uL for
> both construct & transposase.  Does this seem like a reasonable
> concentration?  I saw on another posting that Dr. Lister described
> using ng/uL concntrations.
> Thoughts?
>
> Many thanks!
>
> Jonathan Schumacher
> Research and Development Scientist
> Anatomic Pathology Molecular Diagnostics
> ARUP Institute for Clinical & Experimental Pathology
> 500 Chipeta Way
> Salt Lake City, UT 84108
> Telephone: 801-583-2787 extension 2958
> Fax: 801-584-5109
> Email: schumaj from aruplab.com
>
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