DNA-seq of swabs used on an Apis mellifera Florida swarm trap
DNA-Seq libraries for Illumina NovaSeq sequencing were constructed at the UF-ICBR Gene Expression Core (https://biotech.ufl.edu/gene-expression-genotyping/, RRID:SCR_019145) using the NEBNext UltraTMII FS DNA Library Prep Kit for Illumina (Cat# E7805S). Briefly, DNA samples were fragmented, and end repaired according to the user guide followed by adaptor ligation. Next, the adaptor-ligated DNA were purified by Mag-Bind TotalPure NGS (from Omega Bio-Tek from Omega Bio-Tek, catalog # M1378-02). Finally, The PCR was done for 3-6 cycles of amplification according to the input of DNA. Barcoding was done using the indexing reagents provided in the NEBNext Unique Dual Index Oligos kit (Cat# E6440S). The final libraries were purified by Mag-Bind TotalPure NGS (from Omega Bio-Tek from Omega Bio-Tek, catalog # M1378-02), quantified (QUBIT, ThermoFisher), and sized on the Agilent TapeStation (DNA5000 Screen Tape). The individual libraries were pooled equimolarly and sequenced on the Illumina NovaSeq X Plus using a 10B flow cell (2  150 cycles). The final library loading concentration was 130 pM with 5% PhiX spike-in control. FastQ files were generated using the BCL2fastQ function in the Illumina BaseSpace portal. Sequencing was performed at the UF-ICBR NextGen DNA Sequencing core (https://biotech.ufl.edu/next-gen-dna/, RRID:SCR_019152).