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 have any advice on how to acquire the information on how to get this Transposase mRNA? Do you order it online, do you need to construct it yourself?
If you have any papers on this topic, I'd be more than grateful! Thank you
Dear Monika,
Here you can get all the info you need:
http://tol2kit.genetics.utah.edu/index.php/Main_Page
You need to get the plasmid with cDNA of the Transposase + promoter for transcription. Then you have to use an in vitro transcription system to get your TPAse RNA, check quality and inject. I usued to start by injecting 20-30 pg of each construct + TPAse RNA together.
I hope this helps,
Best regards,
JEsus
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