ena-EXPERIMENT-Carlsberg Research Laboratory-15-12-2017-05:42:05:687-3
PacBio RS paired end sequencing
-
D08_1
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).
Carlsberg Research Laboratory
ena-EXPERIMENT-Carlsberg Research Laboratory-15-12-2017-05:42:05:687-3