Need Sterile Technique for Fruit Tissue Storage - (Feb/27/2013 )
I would like to cut open and snap-freeze some tissue from a cacao fruit (seeds and pericarp). I plan to do HPLC analysis on various tissues, germinate some of the seeds, and perform RNA extractions.
I'd like to know if anyone has suggestions as to what techniques to use so I don't ruin my RNA or secondary compounds as potential analytes (i.e. how to sterilize my blade to eliminate RNases, chemicals to use, etc.)
All help is appreaciated!
Scalpel blades straight out of the packet should be sterile enough. RNAlater might be useful for you, though snap-freezing and storing at -80 is the best option. If you can, work in a laminar flow or class 2 hood to keep your stuff as sterile as possible once you open it. You will need to sterilize the fruit before you put it into the hood, this can be done by wiping in RNase inhibitors such as RNase away and then wiping down with 70% ethanol.
When you say to sterilize the fruit, you do mean to sterilize the outside of the fruit, correct?
yes, sorry should have been a bit more clear there.
No problem Bob, thanks!