RNA-Seq of Homo sapiens : human periodontal ligament stem cells
A total amount of 2 g RNA per sample was used as input material for the RNAsamplepreparations. Sequencing libraries were generated using VAHTS mRNA-seq v2 Library PrepKit for Illumina following manufacturers recommendations and index codes were added toattribute sequences to each sample. Briefly, mRNA was purified from total RNAusingpoly-T oligo-attached magnetic beads. Fragmentation was carried out using fragmentationbuffer. First strand cDNA was synthesized and second strand cDNA synthesis wassubsequently performed. Remaining overhangs were converted into blunt ends. Afteradenylation of 3 ends of DNA fragments, adaptor with hairpin loop structure were ligated. Then the PCR was performed. At last, Qubit HS quantification, Agilent 2100Bioanalyzer/Fragment Analyzer 5300 quality control, the final library size of about 350bp.