In vitro transcription of a large mRNA

Derek Milner dm690785 at mbcr.bcm.tmc.edu
Mon May 2 00:59:17 EST 1994


I am about to embark upon a project designed to determine if a neuronal mRNA
contains a "zipcode" sequence which specifies its subcellular localization. One
of the experiments I plan to do is to synthesize the mRNA in vitro, label by
fluorophore conjugation, microinject into cultured neurons and observe transport
of the mRNA by laser scanning confocal microscopy. My question is this: The
transcript is rather large ( approx 9kbp). Can a transcript of this size be
synthesized by in vitro transcription? I have seen a reference in which a 
viral RNA of slightly larger than 7 kb has been synthesized by in vitro
transcription. Does anyone know if there is a limit to the size of an RNA that
can be synthesized by in vitro transcription, and does anyone have any hints or
advice to give concerning in vitro transcription of large RNAs?

Thanks,

-Derek



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