7a
7a_VO_12wk
RNA-Seq libraries were constructed with the mRNA Sequencing Sample Preparation Kit (Illumina). PolyA+ messenger RNA was selected from 1 ?g of RNA with magnetic oligo (dT) beads, chemically fragmented and converted to cDNA with random hexamers. Double stranded cDNAs were end-repaired, and the 3-ends were A-tailed followed by ligation of Illumina sequencing and amplification adapters randomly to the ends. The adaptor-ligated cDNAs were loaded onto 2% agarose E-gels (Invitrogen) and the fraction containing 200500 bp was excised. Size-selected cDNAs were amplified by PCR with primers that introduced unique barcodes to each library. The libraries were then quantitated with the Qubit (Life Technologies) and the average size was determined on an Agilent bioanalyzer DNA7500 DNA chip (Agilent Technologies) and diluted to 10nM. Each library was loaded onto one lane of an 8-lane flowcell for cluster formation and sequenced on an Illumina Genome Analyzer IIx according to the manufacturers protocols. The fastq files were generated with Casava version 1.6.
7a
SUB7860719
7a