title: PacBio RS paired end sequencing
design: -
library_name: C10_1
library_construction_protocol: A total of 5μg for each sample was used as input into library preparation. The SMRTbell library was constructed with SMRTbell™ Template Prep Kit 1.0 (PN 100-259-100) following manufacture's instructions (Pacific Biosciences). The small fragments lower than 20 kb of SMRTbell template was removed using Blue Pippin Size selection system for large-insert library. The constructed library was validated by Agilent 2100 Bioanalyzer. After a sequencing primer is annealed to the SMRTbell template, DNA polymerase is bound to the complex using DNA/Polymerase Binding kit P6. This polymerase-SMRTbell-adaptor complex is then loaded into SMRT cells. The SMRTbell library was sequenced using 4 SMRT cells (Pacific Biosciences) using C4 chemistry (DNA sequencing Reagent 4.0) and 240 minute movies were captured for each SMRT cell using the PacBio RS II (Pacific Biosciences) sequencing platform(1).