Relationship Between Tree Frog And Bromeliad [NEW]
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Relationship Between Tree Frog And Bromeliad [NEW]
The extracted DNA was stored in 1.5 ml cryogenic carriers at -20°C. After aerosolization, each sample was split into three carrier volumes (75, 30 and 15 ml). Each carrier was labeled with sample data and placed in a single box, which was airlifted to California for filtering on site at CIDS and storage at -78C. All sequencing reactions were performed at the ZMBH in Germany or the NYBG in Puerto Rico.
By combining three samples and aerosolizing that volume, we were able to use 95% less sample when compared to a single sample from an entire tree. This method was necessary to collect representatives of P. auratus while minimizing the number of frogs and bromeliads that were disturbed. In a separate study, we demonstrated that P. auratus could be detected using a hubba filter (10 micron nylon mesh, Sigma)
We attempted to amplify P. auratus DNA from the original unfiltered bromeliad water samples. We failed to amplify the 18S rRNA gene in all 29 samples; however, we recovered an COI fragment that was 99% identical to that obtained from prototype C-2 from the same area in the US. These results suggest that copepods found in bromeliad water or the unlikely possibility that DNA was transferred from the frog eardrum or mouth. d2c66b5586