GSM7679418: Ant. No. 38 , Size: 6348.66 µM, miRNA-seq; Camponotus floridanus; miRNA-Seq
DNA and RNA (including miRNA) were extracted with the AllPrep DNA/RNA/miRNA Universal Kit (QIAGEN) according to the manufacturer's instructions. Total RNA was quantified using a NanoDrop Spectrophotometer ND-1000 (NanoDrop Technologies, Inc.) and its integrity was assessed using a 2100 Bioanalyzer (Agilent Technologies). Libraries were generated from 1000 ng of total RNA using the TruSeq® Small RNA Sample Prep Kit (Illumina), as per the manufacturer's recommendations. cDNA construct purification has been performed on a Pippin Prep instrument (SAGE Science). Final libraries were quantified using the Kapa Illumina GA with Revised Primers-SYBR Fast Universal kit (Kapa Biosystems). Average size fragment was determined using a LabChip GX (PerkinElmer) instrument. The libraries were normalized and pooled at 3nM 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 now at 200pM was loaded on a Illumina cBot and the flowcell was ran on a HiSeq 4000 for 1x50 cycles. A phiX library was used as a control and mixed with libraries at 1% level.