| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
Total RNA of the 10 samples were mixed together in equal parts (2 µg for each sample). The total RNA was reverse transcribed using the Clontech SMARTerTM PCR cDNA synthesis Kit with anchored oligo(dT)30 as the primer. The double-strand cDNA was then amplified with LD-PCR (Long-Distance PCR) using Advantage 2 PCR Kit. We used the BluePinppin size selection system (Sage Science, Beverly, MA) to generate cDNA fractions with different sizes including 0.5-1Kb, 1-2 Kb, 2-3 Kb and >3 Kb. These libraries were then constructed with the Pacific Biosciences’ SMRTbell Template Prep Kit 1.0, following the manufacturer’s protocol. A total of six SMRT cells (2 SMRT cell for libraries of 1-2 Kb; 2 SMRT cell for libraries of > 3Kb; one SMRT cell for libraries of 0.5-1 Kb, and; one SMRT cell for libraries of 2-4 Kb) were sequenced on the PacBio RS II platform. |
FL-cDNA |
TRANSCRIPTOMIC |
unspecified |
SINGLE
|
|
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