## 5 Materials and Methods  
_A. domesticus_ colony maintenance. _A domesticus_ were originally purchased from a US insect farm in 2018 and maintained as a lab colony for multiple generations. They were feda diet of specially formulated cricket feed from LoneStar (TFP Nutrition) (Nacogdoches, TX, USA) ([[http://lonestarfeed.com](http://lonestarfeed.com)]([http://lonestarfeed.com](http://lonestarfeed.com))), which was the same feed used by the insect farm. Feed and water were provided to each cricket cage twice weekly. Eggs were collected twice per week from wild type adults in a petri dish (150 mm diameter \(\times\) 10 mm deep ) filled with hydrated (deionized water saturated) polyacrylamide water crystals, as crickets prefer to lay eggs in moist areas. Each week, the old egg-lay dish was replaced with a new dish on Monday and Friday. Eggs were collected each Tuesday for colony maintenance. For egg collection, the petri dish with water crystals containing eggs was washed into a glass beaker using deionized water. Additional water was added to the beaker and stirred a few times, and eggs were allowed to settle to the bottom for approximately 10-15 s. Most of the excess water and crystals were slowly removed to retain eggs. Washing was repeated 2-3 times, and a plastic Pasteur pipette was used to aspirate eggs from the bottom of the beaker and transfer to a piece of damp/moist paper towel in a petri dish lid. The lid was placed into an 8 oz Deli container (S-21216, Ulime, Pleasant Prairie, WI, USA) with a lid until the eggs hatched (approximately 8-10 days). After hatching, crickets were transferred into cricket cages. Cricket cages consisted of 20 qt plastic storage containers (Gasket Box, Sterilite, Townsend, MA, USA) with four 100 mm circles cut on the side and two on the lid that were covered by screen mesh material. Cytometric genome size estimation of _A. domesticus_. The 1C (haploid) genome size of males and females of _A. domesticus_ was estimated as described in [PERSON] et al. (2019)[49]. In brief, a single _A. domesticus_ head, a single _female_ head (1C = 328 Mbp), and a portion of brain from a _Periplaneta americana_ male (1C = 3300) were placed together into 1 mL of ice-cold Galbraith buffer in a 2 mL Dounce tissue grinder. Nuclei were released by grinding with 10 strokes of an A (loose) pestle, then filtered through nylon mesh into a 1.5 mL microfuge tube. The DNA in the released nuclei were stained by adding 25 \(\mu\)L of propidium iodide (1 mg/mL) and allowed to stain 3 h in the dark at 4 \({}^{\circ}\)C. The mean red PI fluorescence of the stained DNA in the nuclei of the sample and the standards was quantified using a CytoFlex flow cytometer (Beckman Coulter). Haploid (1C) DNA quantity was calculated as (2C sample mean fluorescence/2C standard mean fluorescence) times 328 Mbp for the _D. virilis_ standard and times 3300 Mbp for the _P. americana_ standard. The estimates based on the two standards were averaged to produce a 1C estimate for each sample. Genome sequencing and assembly. A single adult male _A. domesticus_ was shipped to Dovetail Genomics (now Cantata Bio, Scotts Valley, CA, USA). Genomic DNA was extracted and shipped to North Carolina State University and USDA ARS, Stoneville, MS for long read sequencing, and extracted nuclei were used for Chicago and Hi-C libraries. Long read data was obtained using Pacific Biosciences technologies (PacBio, Menlo Park, CA, USA). Libraries were prepared using SMRTbell Express Template Prep Kit, and Sequel Binding Kit 2.1 was used for polymerase binding. The binding reaction was loaded at 8 pmol concentration with a 15 h run time. The gDNA was sequenced on eight LR SMRTCells 1M v3 for the Sequel I for a total of 58.7 Gb of long read data. The genome was assembled from PacBio reads using CANU v1.8[88]. The two sets of PacBio CLR reads were corrected separately and co-assembled.