Dieffenbachia Biforine Transcriptome
A transcriptome is the sum total of all the genes being transcribed
in a particular organism, tissue, or cell. We would like to determine
what transcripts are being made in Dieffenbachia biforine idioblasts
and to compare this to the transcripts being made in mesophyll cells.
The ultimate goal is to clone these transcripts as cDNAs and to sequence
them to determine, if possible, what genes they represent.
we have to get RNA out of a small quantity of isolated cells, a daunting
task. To make matters worse, oxalate, a key ingredient of those
crystals, interferes with RNA extraction. A search for
Dieffenbachia RNA in the literature turned up exactly zero papers,
suggesting no one has tried getting RNA from this species before.
Drummond Vogan was the first to try, working out the quirks, but finding
nothing. Jeremy Hensley bravely tried, and got a little RNA out of
whole leaf, but nothing out of cells. Brad Howard continued the work
with more frustration and just as little result. At that point we took
a step back and tried several available methods for RNA extraction from
tough plant tissues. By the end of spring semester 2006 Brad had
excellent RNA extracts from Dieffenbachia leaf. Currently, Brian Prall
is accumulating deep frozen idioblast cells to test Brad's method on.
At left, Brad superchills his equipment with liquid nitrogen from a beer mug
before beginning an RNA extraction.