[Coral-List] Comparison of extraction kits to obtain bacterial DNA from coral tissue samples
ckellogg at usgs.gov
Fri May 23 09:32:07 EDT 2014
Many of us have struggled with coral samples, and the only thing that is
consistent is that nothing works the same for every coral species! Last
summer, one of my students compared three MO BIO kits (PowerPlant - now
discontinued, PowerPlantPro - the replacement, and PowerSoil) using the
modifications outlined in Sunagawa et al. 2010 [PLOS ONE 5(3): e9554.
doi:10.1371/journal.pone.0009554] to see which worked best. She tested
across four different coral species: *Orbicella annularis*, *Siderastrea
siderea*, *Lophelia pertusa* and *Primnoa resedaeformis*.
Coral microbiology is an expanding field, yet there is no standard DNA
extraction protocol. Although many researchers depend on commercial
extraction kits, no specific kit has been optimized for use with coral
samples. Both soil and plant DNA extraction kits from MO BIO Laboratories,
Inc., have been used by many research groups for this purpose. MO BIO
recently replaced their PowerPlant® kit with an improved PowerPlantPro kit,
but it was unclear how these changes would affect the kit’s use with coral
samples. In order to determine which kit produced the best results, we
conducted a comparison between the original PowerPlant kit, the new
PowerPlantPro kit, and an alternative kit, PowerSoil, using samples from
several different coral genera. The PowerPlantPro kit had the highest DNA
yields, but the lack of 16S rRNA gene amplification in many samples
suggests that much of the yield may be coral DNA rather than microbial DNA.
The most consistent positive amplifications came from the PowerSoil kit.
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Christina A. Kellogg, Ph.D.
St. Petersburg Coastal and Marine Science Center
US Geological Survey
600 4th Street South
St. Petersburg, Florida 33701
PH: (727) 502-8128 ***Note new phone and fax numbers***
FAX:(727) 502-8181 ***As of Nov 19, 2013***
Email: ckellogg at usgs.gov
The person who says it cannot be done should not interrupt the person
doing it. -- Chinese proverb
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