title: GSM7677552: Ant No. 44 , Size: 7114.6 µM, WGBS; Camponotus floridanus; Bisulfite-Seq
library_construction_protocol: DNA and RNA (including miRNA) were extracted with the AllPrep DNA/RNA/miRNA Universal Kit (QIAGEN) according to the manufacturer's instructions. gDNA was quantified using the Quant-iT™ PicoGreen® dsDNA Assay Kit (Life Technologies). Libraries were generated from 250ng of gDNA using the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England BioLabs).  Adapters were purchased from NEB. Size selection of libraries containing the desired insert size was performed using sparQ PureMag Beads (Quantabio). Bisulfite conversion had been carried out with the EZ DNA Methylation-Lightning Kit (Zymo). Libraries were quantified using the KAPA Library Quanitification Kits - Complete kit (Universal) (Kapa Biosystems). Average size fragment was determined using a LabChip GX (PerkinElmer) instrument. The libraries were normalized and pooled and then denatured in 0.05N NaOH and neutralized using HT1 buffer. ExAMP was added to the mix following the manufacturer's instructions. The pool was loaded at 250pM on an Illumina cBot and the flowcell was run on a HiSeq X for 2x151 cycles (paired-end mode). A phiX library was used as a control and mixed with libraries at 1% level.