Collection: Bulk zooplankton were collected at an open ocean time series site in the North Pacific Subtropical Gyre (station ALOHA, 22.45N, 158W) during 11 of the routine Hawai'i Ocean Time-series (HOT) research cruises at approximately monthly intervals from September of 2012 to October of 2013 (HOT-246 to HOT-256). Mesozooplankton were collected using a 1 m2, 200 um-mesh ring net towed obliquely from a mean maximum depth of 155 m (SD = 31 m) to the sea surface. Zooplankton for this study were collected from three nighttime tows completed between the hours of 2200-0200 on consecutive nights for each sampling period so that all collections for a single cruise were collected within three days of one another. Following net retrieval, bulk plankton were quantitatively split using a Folsom plankton splitter, and 1/4 of the material was preserved in 95% non-denatured ethyl alcohol (EtOH) and stored at -20C. From these bulk collections, 50 Haloptilus longicornis and 50 Pleuromamma xiphias individuals were sorted from each net tow for use in this study.

mtDNA Sequence Generation: For both H. longicornis and P. xiphias, DNA was extracted using the Qiagen DNeasy Blood and Tissue Kit (Qiagen, Inc., Valencia, CA). For H. longicornis, a 546 base-pair fragment of the mitochondrial cytochrome c oxidase subunit II (mtCOII) gene was amplified by polymerase chain reaction (PCR) with species-specific primers COII_F6 and COII_R9. For P. xiphias, a 551 base-pair fragment of the mitochondrial cytochrome c oxidase subunit I (mtCOI) was amplified by PCR using species-specific primers PLXI_VH and PLXI_VL. Specific information on primer sequences, PCR reaction mixes, thermal cycler conditions, and PCR purification is provided in the manuscript associated with this submission. Purified PCR products were sequenced on an ABI 3730XL capillary sequencer (Applied Biosystems, Foster City, CA). Sequences were aligned, edited and trimmed using Geneious 7.0.6 (Biomatters, Ltd., Auckland, New Zealand). Unique haplotypes were identified using the Haplotype Collapser and Converter in FaBox 1.35 (http://users-birc.au.dk/biopv/php/fabox/), and deposited with their respective protein translations in GenBank under accession numbers: KY560470 - KY560565. An mtDNA sequence for each individual specimen is deposited in this BCO-DMO submission as part of one of two fasta files. Each fasta file contains the mtDNA sequence fragments from all individuals from a single species aligned together.

Methodology is further described in the paper itself:
Iacchei, M., E. Butcher, E. Portner, Goetze, E. (in press) It’s about time: Insights into temporal genetic patterns in oceanic zooplankton from biodiversity indices. Limnology and Oceanography