A Method for Fixation of Fish Larvae for Morphological and Genetic Studies
- 12 Downloads
Three aqueous buffer solutions that make it possible to stabilize DNA and also preserve the initial body shape and morphological structures are tested for fixation and preservation of deep-sea fishes’ larvae. Based on the quality assessment of DNA and appearance of the larvae 6 months after fixation, the buffer solution containing dimethyl sulfoxide is recommended for use. Unlike ethanol or formalin, this fixative is nontoxic, nonflammable, does not require storage of samples in the refrigerator, and is also compatible with standard DNA extraction methods or commercial kit protocols.
Keywords:fish larvae fixatives dimethyl sulfoxide–containing buffer solution DNA barcoding mtDNA cox 1
This study was supported in part by the Russian Science Foundation, project no. 19-14-00026 (expedition works), the Russian Foundation for Basic Research, project no. 18-04-00019, and within the framework of state assignment no. 0112-2018-0002 (laboratory analysis).
- 8.Hykin, S.M., Bi, K., and McGuire, J.A., Fixing formalin: a method to recover genomic-scale DNA sequence data from formalin-fixed museum specimens using high-throughput sequencing, PLoS One, 2015, vol. 10, no. 10, p. e0141579. https://doi.org/10.1371/journal.pone.0141579
- 13.Longmire, J.L., Maltbie, M., and Baker, R.J., Use of “lysis buffer” in DNA isolation and its implications for museum collections, Occas. Pap. Mus. Texas Tech. Univ., 1997, vol. 163, pp. 1–3.Google Scholar
- 14.Maniatis, T., Fritsch, E.F., and Sambrook, J., Molecular Cloning, a Laboratory Manual, New York: Cold Spring Harbor Lab., 1982.Google Scholar
- 16.NCBI, National Center for Biotechnology Information, 2018. https://www.ncbi.nlm.nih.gov/Blast.cgi.Google Scholar
- 17.Quach, N., Goodman, M.F., and Shibata, D., In vitro mutation artifacts after formalin fixation and error prone translesion synthesis during PCR, BMC Clin. Pathol., 2004, vol. 4, no. 1. https://doi.org/10.1186/1472-6890-4-1
- 20.Pisani, G.R., A Guide to Preservation Techniques for Amphibians and Reptiles, Lawrence: Soc. Study Amphibians Reptiles, 1973.Google Scholar
- 30.Ward, R.D., Zemlak, T.S., Innes, B.H., et al., DNA barcoding Australia’s fish species, Philos. Trans. R. Soc., B, 2005, vol. 360, pp. 1847–1857.Google Scholar