title: GSM2258365: 6-days hypocotyl, biological replicate 1; Cannabis sativa; RNA-Seq
library_construction_protocol: Libraries were prepared from 3µg of total RNA using the SMARTer Stranded RNA-Seq kit (Clontech). The isolation of mRNAs was performed using the Illumina beads and the TruSeq protocol (Illumina).The final elution of the mRNAs was performed using the Illumina elution buffer (19.5 µl).  The isolated mRNAs were quantified using a Qubit fluorometer, as described above. 10 ng mRNA were used for the cDNA synthesis and shearing. Indexing was performed using the Illumina indices 1-12. The enrichment step was carried out using 12 cycles of PCR. Subsequently, libraries were checked using 2100 Bioanalyzer (DNA-1000) to evaluate the mean fragment size. Quantification was done using the KAPA library quantification kit (KAPA Biosystems) using a ViiA7 Real-Time PCR System (Applied Biosystems). The pooled libraries were sequenced on an Illumina MiSeq in 6 consecutive runs (2x150 paired end: MiSeq reagent kit V3, 300cycles).