October 1, 2017.
A coastwide bloom of the toxigenic diatom Pseudo-nitzschia in 2015 resulted in the largest reCryptoheros septemfasciatus is a species of cichlid fish (family Cichlidae) that varies phenotypically from population to population. We do not know the relatedness of the different populations of C. septemfasciatus in the many geographically close rivers of northern Costa Rica. To determine this, we will ultimately need to collect many fish and/or tissues at many sites in the wild. It is critical that we know whether we are able to obtain DNA from various tissues (e.g fin clips) and/or from fish fry. I assisted in creating and perfecting a protocol for DNA analysis using fish fry due to the fact that fry are much easier to capture and transport than adults. I asked: (1) do cichlid fry have DNA that we are able to sequence for analysis; (2) if so, at what size fry is that DNA evident; and (3) can we obtain DNA from freshly dead, frozen, and preserved fishes? To answer these questions, I captured and preserved 45 cichlid fry and 7 fin clips from multiple fishes. I extracted DNA from all the samples using Qiagen DNA Extraction Kits, then conducted polymerase chain reactions (PCR) for each using the primers Acit1F and Acit1R. I ran Gel Electrophoresis’ to check for DNA amplification. I was able to amplify DNA in 25 fry and 4 fin clips. This answered the questions that DNA can be found in cichlid fry as small as 5.2mm and that DNA is still available for use from fishes that are frozen and preserved in formalin. Although the primers I used were unable to copy and amplify DNA in all 45 of the fry, I was able to amplify DN. Overall, we now know that cichlid fry do have DNA available for analysis as do frozen fishes and fishes preserved in formalin.
California State University, Sacramento (Sac State)
This material is based upon work supported by the National Science Foundation through the Robert Noyce Teacher Scholarship Program under Grant # 1136419. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation. The research was also made possible by the California State University STEM Teacher and Researcher Program, in partnership with Chevron (www.chevron.com), the National Marine Sanctuary Foundation (www.marinesanctuary.org) and Sacramento State University.