Authors: Avery Larsen, Heath Ogden
Mentors: Heath Ogden
Insitution: Utah Valley University
Mayflies, also known as Ephemeroptera, are members of the anciently derived infraclass known as Paleoptera, the first group of insects that evolved the ability to fly (Gillott, 2005). Distinct developmental characteristics of the life stages of the mayfly are the reason that ephemeroptera are of particular interest. The life stages are; the egg, nymphs, subimago, and imago. The aquatic nymph stage does not have wings but instead has leaf-shaped gills that can be used to help propel the insect through its environment (Eastham, 1936). Centroptilum triangulifer will be used for RNA extractions to study the development of both wings and gills. Objectives for this research are 1). Elucidate, describe, capture, and record distinguishing characteristics of the different instars of Centroptilum triangulifer. 2) Identify key instars integral to gill and wing development in Centroptilum triangulifer. 3) Perform 10 RNA extractions. 4) Use Qubit technology to ensure ≥ 20 ng/μL RNA concentration per ≥ 10 μL. 5) samples will be packaged in thermo-stable shipping boxes and sent to Novogene where samples will be tested once more for quality control. 7) Once the quality is confirmed, samples will be processed using poly-A enrichment and then sequenced using high-throughput Illumina sequencing. Data will be tested for quality control and then sent to Ogden labs. 8). Using an in-house bioinformatics workflow, RNA data will be checked for quality, trimmed, and aligned, before RNA data is first trimmed, and then aligned to Ogden labs transcriptome reference genome. After alignment, the number of reads per gene, or hit counts, are calculated and compared. 9) RNA results of different instars will be compared to each other as well as NCBI databases using the Basic Local Alignment Search Tool.