TECHNICAL BREAKTHROUGHS

E. chlorotica produces and secretes copious amounts of muco- polysaccharides, which interfere with almost all attempts to analyze these animals at the sub-cellular level when employing standard protocols.  Much of our earlier effort was spent on developing techniques to overcome the mucous problem.  This involved approaches as simple as rolling the sea slugs gently in Kimwipes to exude the mucous to developing extensive purification protocols for nucleic acids and proteins.  An early breakthrough was the discovery that a mucolytic agent, N-acetyl-L-cysteine (mucomyst), was effective in breaking up the mucous when added to the homogenization media.  However, one has to be aware that this compound is also a DNase.  We are continuing to improve upon the experimental techniques to decrease the number of steps required for each purification.  Most recently we have found that some of the new commercial kits and reagents produced by Molecular Research Center, Inc., can be used with the sea slugs.  For example, extra-strength DNAzol is an excellent reagent for quickly isolating DNA from the sea slugs in very high yields and, although TRIzol was not as effective in isolating DNA, it did yield RNA useable for Northern analysis.