Xestocephalus

Transcriptome of Xestocephalus desertorum

"Construction of libraries and sequencing on the Illumina HiSeq2500 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 HiSeq2500 with TruSeq SBS sequencing kits version 4. The libraries were sequenced from both ends of the molecules to a total read length of 100nt from each end. The raw .bcl files were converted into demultiplexed fastq files with Casava 1.8.2 (Illumina)."

{'breed': 'wildtype', 'tissue': 'whole insect', 'lat_lon': '40.090439 N 87.836511 W', 'collected_by': 'Christopher H. Dietrich', 'geo_loc_name': 'USA: IL, Vermillion Co., 1.5 mi SSE Muncie', 'biosamplemodel': 'Invertebrate', 'collection_date': '2013-07-31', 'isolation_source': 'wild-caught', 'specimen_voucher': 'P070'}

RNA-seq of Xestocephalus sp.

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': '2110 m', 'collected_by': 'Hu Yanhua', 'geo_loc_name': 'China:Shaanxi', 'identified_by': 'Dai Wu', 'biosamplemodel': 'Invertebrate', 'collection_date': '2017-06-22', 'isolation_source': 'wild-caught'}