Sitodiplosis mosellana

RNA seq of sitodiplosis mosellana adult male

For Illumina RNA-Seq, the library was prepared by NEBNext Ultra RNA Library Prep Kit. Briefly, the enriched mRNA was purified using Oligo (dT) magnetic beads. Subsequently, the mRNA was processed into short fragments by fragmentation buffer, and a strand of cDNA was synthesized by random hexamers using the mRNA as the template. Then AMPure XP beads were used to purify the two-strand cDNA synthesized by buffer, DNA polymerase I and dNTPs. Finally, AMPure XP beads were used to select 250300 bp fragments, and the final cDNA library was obtained through PCR enrichment. Qubit2.0 Flurometer (Life Technologies,CA, USA) was used to perform preliminary quantification of the constructed library, and the library was diluted to 1 ng / L. Then Agilent 2100 was adopted to detect the insert size length of the library, and the Q-PCR kit was used to accurately quantify the effective concentration of the library (effective library concentration>2 nM) to ensure the quality of the library.

{'host': 'wheat', 'tissue': 'male adult', 'isolate': 'Zhongjun Gong', 'geo_loc_name': 'China: Puyang', 'biosamplemodel': 'Invertebrate', 'collection_date': '2019-04-12', 'isolation_source': 'soil replicate 1'}

RNA seq of sitodiplosis mosellana adult male

For Illumina RNA-Seq, the library was prepared by NEBNext Ultra RNA Library Prep Kit. Briefly, the enriched mRNA was purified using Oligo (dT) magnetic beads. Subsequently, the mRNA was processed into short fragments by fragmentation buffer, and a strand of cDNA was synthesized by random hexamers using the mRNA as the template. Then AMPure XP beads were used to purify the two-strand cDNA synthesized by buffer, DNA polymerase I and dNTPs. Finally, AMPure XP beads were used to select 250300 bp fragments, and the final cDNA library was obtained through PCR enrichment. Qubit2.0 Flurometer (Life Technologies,CA, USA) was used to perform preliminary quantification of the constructed library, and the library was diluted to 1 ng / L. Then Agilent 2100 was adopted to detect the insert size length of the library, and the Q-PCR kit was used to accurately quantify the effective concentration of the library (effective library concentration>2 nM) to ensure the quality of the library.

{'host': 'wheat', 'tissue': 'male adult', 'isolate': 'Zhongjun Gong', 'geo_loc_name': 'China: Puyang', 'biosamplemodel': 'Invertebrate', 'collection_date': '2019-04-12', 'isolation_source': 'soilreplicate 2'}

PacBio Isoseq of Sitodiplosis mosellana: pooled adults and pooled 3rd instar larvae

Total RNA was extracted from either 100 3rd instar larvae or 20-30 adults using the Cytiva RNAspin Mini kit. Each was quantified on a nanodrop spectrophotometer, then pooled in equal amounts (equal nanograms) prior to library generation. Two libraries were generated using : a library with standard size selection and another with selecting long reads. These libraries were pooled 80% standard :20% long reads and sequenced (without barcoding) on the same chip.

{'age': '3rd instar and adult', 'sex': 'mixed', 'tissue': 'whole insects', 'isolate': 'pooled adults and 3rd instar larvae', 'geo_loc_name': 'Canada: Edmonton', 'biosamplemodel': 'Invertebrate', 'collection_date': '2021', 'isolation_source': 'University of Alberta colony manitained by Dr. Boyd Morii'}