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Fig. 1 | BMC Medical Genomics

Fig. 1

From: Biomarker discovery: quantification of microRNAs and other small non-coding RNAs using next generation sequencing

Fig. 1

Illustration of study design and samples. Human biological samples (N = 45) were included in the present study. a Peripheral blood from a single individual was split into 11 aliquots (technical replicates) to test three different small RNA library purification methods: Novex TBE PAGE gel (N = 3), Pippin Prep automated gel system (PPS) (N = 4), and AMPure XP beads ((N = 3). Sample C1 (control-human brain) (N = 1), sample AC (control-no purification method) (N = 1). b Peripheral blood from a single individual was split into 5 aliquots (technical replicates) to test optimal amounts of RNA input: (1 μg), (0.5 μg), (0.25 μg), (0.1 μg), and (0.05 μg). All libraries were purified using the PPS system. c Peripheral blood samples from 15 healthy volunteers (biological replicates) to test the effects of RNA integrity. Samples were split into 5 groups (N = 3) with average RIN values of 9, 7, 5.4, 2.2 and 0. All libraries were purified using AMPure XP beads. d Peripheral blood samples from 12 healthy volunteers (biological replicates) to test effects of sequencing coverage. Samples sequenced on both a HiSeq2500 (N = 12) and MiSeq (N = 12) Illumina sequencers. All libraries were purified using AMPure XP beads. e Human whole-blood (N = 4), brain (N = 4), heart (N = 4) and liver (N = 4) tissues to test expression and tissue specificity of small ncRNAs. All libraries were purified using AMPure XP beads

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