Psylliodes chrysocephalus

RNA degradome-seq: Psylliodes chrysocephala: 10d-adult

We isolated total RNA from 20 adults using the Quick-RNA Tissue kit (Zymo Research). Polyadenylated RNA was purified using Dynabeads mRNA Purification kit (ThermoFisher) and Oligo d(T) 25 Magnetic Beads (NEB). The 5 RNA adaptor (100 M, Tab. S2) was ligated to RNA using T4 RNA ligase buffer (NEB) and 5 RNA adaptor ligated polyadenylated RNA was purified using Dynabeads mRNA purification kit (ThermoFisher). was First cDNA strand was synthesized using ProtoScript TM II reverse transcriptase (NEB). The reaction for the second strand cDNA synthesis included first cDNA strand, MyFi Mix (BioCat) and 5 and 3 adaptor primers (Tab. S2) and 15 PCR cycles were performed. The PCR product was purified using Monarch PCR & DNA cleanup kit and digested with EcoP151 (NEB). dsDNA adaptors (Tab. S2) were ligated to the digested DNA using T4 DNA ligase (NEB). The dsDNA adaptor ligated DNA (79 bp) was purified through PAGE and concentrated through ethanol precipitation. Thirteen PCR cycles were performed using MyFi Mix (BioCat) to add the SR1 indexes (Tab. S2) to the libraries. The libraries were pooled and purified through PAGE.

{'age': '10 day old', 'sex': 'female', 'host': 'Brassica napus subsp. napus', 'tissue': 'whole body', 'isolate': 'lab', 'geo_loc_name': 'Germany', 'biosamplemodel': 'Invertebrate', 'collection_date': '2024'}

RNA degradome-seq: Psylliodes chrysocephala: 15d-adult

We isolated total RNA from 20 adults using the Quick-RNA Tissue kit (Zymo Research). Polyadenylated RNA was purified using Dynabeads mRNA Purification kit (ThermoFisher) and Oligo d(T) 25 Magnetic Beads (NEB). The 5 RNA adaptor (100 M, Tab. S2) was ligated to RNA using T4 RNA ligase buffer (NEB) and 5 RNA adaptor ligated polyadenylated RNA was purified using Dynabeads mRNA purification kit (ThermoFisher). was First cDNA strand was synthesized using ProtoScript TM II reverse transcriptase (NEB). The reaction for the second strand cDNA synthesis included first cDNA strand, MyFi Mix (BioCat) and 5 and 3 adaptor primers (Tab. S2) and 15 PCR cycles were performed. The PCR product was purified using Monarch PCR & DNA cleanup kit and digested with EcoP151 (NEB). dsDNA adaptors (Tab. S2) were ligated to the digested DNA using T4 DNA ligase (NEB). The dsDNA adaptor ligated DNA (79 bp) was purified through PAGE and concentrated through ethanol precipitation. Thirteen PCR cycles were performed using MyFi Mix (BioCat) to add the SR1 indexes (Tab. S2) to the libraries. The libraries were pooled and purified through PAGE.

{'age': '15 day old', 'sex': 'female', 'host': 'Brassica napus subsp. napus', 'tissue': 'whole body', 'isolate': 'lab', 'geo_loc_name': 'Germany', 'biosamplemodel': 'Invertebrate', 'collection_date': '2024'}

RNA degradome-seq: Psylliodes chrysocephala: 3d-adult: dsProsB1treatment

We isolated total RNA from 20 adults using the Quick-RNA Tissue kit (Zymo Research). Polyadenylated RNA was purified using Dynabeads mRNA Purification kit (ThermoFisher) and Oligo d(T) 25 Magnetic Beads (NEB). The 5 RNA adaptor (100 M, Tab. S2) was ligated to RNA using T4 RNA ligase buffer (NEB) and 5 RNA adaptor ligated polyadenylated RNA was purified using Dynabeads mRNA purification kit (ThermoFisher). was First cDNA strand was synthesized using ProtoScript TM II reverse transcriptase (NEB). The reaction for the second strand cDNA synthesis included first cDNA strand, MyFi Mix (BioCat) and 5 and 3 adaptor primers (Tab. S2) and 15 PCR cycles were performed. The PCR product was purified using Monarch PCR & DNA cleanup kit and digested with EcoP151 (NEB). dsDNA adaptors (Tab. S2) were ligated to the digested DNA using T4 DNA ligase (NEB). The dsDNA adaptor ligated DNA (79 bp) was purified through PAGE and concentrated through ethanol precipitation. Thirteen PCR cycles were performed using MyFi Mix (BioCat) to add the SR1 indexes (Tab. S2) to the libraries. The libraries were pooled and purified through PAGE.

{'age': '3 day old', 'sex': 'female', 'host': 'Brassica napus subsp. napus', 'dsrna': 'dsProsB1', 'tissue': 'whole body', 'isolate': 'lab', 'geo_loc_name': 'Germany', 'biosamplemodel': 'Invertebrate', 'collection_date': '2024'}