Macropsini

Transcriptome of Pedionis garuda

Construction of libraries and sequencing on the Illumina HiSeq4000 were performed at the W. M. Keck Center for Comparative and Functional Genomics at the University of Illinois at Urbana-Champaign. Messenger RNA was selected from one microgram of high quality total RNA. RNAseq libraries were constructed using the TruSeq Stranded RNA Sample Preparation Kit (Illumina, San Diego, CA). Libraries were sequenced on an Illumina HiSeq4000 with HiSeq 4000 sequencing kit version 1. The libraries were sequenced from both ends of the molecules to a total read length of either 100nt or 150nt from each end. The raw .bcl files were converted into demultiplexed fastq files with bcl2fastq v2.17.1.14 Conversion Software (Illumina).

{'breed': 'wildtype', 'tissue': 'whole insect', 'collected_by': 'C. Dietrich', 'geo_loc_name': 'China: Guangxi, Longtan National Forest Park', 'biosamplemodel': 'Invertebrate', 'collection_date': '2017-09-23', 'isolation_source': 'wild-caught'}

RNA-seq of Macropsis matsumurana

After the total RNA extraction and DNase I treatment, magnetic beads with Oligo (dT ) are used to isolate mRNA. Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments. Then cDNA is synthesized using the mRNA fragments as templates. Short fragments are purified and repaired at the end, added single nucleotide A (adenine) and connected with adapters. The suitable fragments are selected for the PCR amplification as templates. After the construction of the library, Qubit2.0 and Agilent 2100 are used to detect the concentration and insert size of the library, and Q-PCR method is used to quantify the effective concentration of the library to ensure the quality of the library. At last, the library is sequenced using Illumina NovaSeq 6000.

{'breed': 'wildtype', 'tissue': 'whole insect without the abdominal genital part', 'altitude': '10 m', 'collected_by': 'Hu Yanhua', 'geo_loc_name': 'China:Zhejiang', 'identified_by': 'Yang Liyuan', 'biosamplemodel': 'Invertebrate', 'collection_date': '2017-05-31', 'isolation_source': 'wild-caught'}

RNA-seq of Pediopsoides kurentsovi

After the total RNA extraction and DNase I treatment, magnetic beads with Oligo (dT ) are used to isolate mRNA. Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments. Then cDNA is synthesized using the mRNA fragments as templates. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters. The suitable fragments are selected for the PCR amplification as templates. During the QC steps, Agilent 2100 Bioanaylzer and ABIStepOnePlus Real-Time PCR System are used in quantification and qualification of the sample library. At last, the library is sequenced using Illumina.

{'breed': 'wildtype', 'tissue': 'whole insect without the abdominal genital part', 'altitude': '1610 m', 'collected_by': 'Hu Yanhua', 'geo_loc_name': 'China:Shaanxi', 'identified_by': 'Yang Liyuan', 'biosamplemodel': 'Invertebrate', 'collection_date': '2017-06-19', 'isolation_source': 'wild-caught'}