proteoglycan removal from extracted RNA

Sara Edge via methods%40net.bio.net (by sedge4 from hboi.fau.edu)
Thu Jul 28 14:59:05 EST 2011


Does anyone know of a method for removing proteoglycan contaminants from
total RNA extracts? I work with marine invertebrates (corals) and
certain species are difficult due to RNA that cannot be successfully
manipulated downstream (i.e. labeling, RT, etc.). Other research have
suggested that endogenous proteoglycans interfere with enzymes that bind
RNA. I use the TRIzol method of extraction and the Qiagen MinElute kit
for purifications. I am getting good quality RNA based on nanodrop
spectrophotometry (260:280 @ 2.0) and Agilent Bioanalyzer analysis
(clean gels, RIN @ 8-9). My 260:230's could be better and tend to be all
over the place, but have gotten up to 1.9. I would appreciate any
suggestions.


Thank you.

 

Sara Edge, Ph.D.

Marine Ecosystem Health

Harbor Branch Oceanographic Institute at FAU

5600 US-1 North, Fort Pierce, FL 34946

772-242-2261

sedge4 from hboi.fau.edu

 



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