title: GSM2254735: MO24-32977011; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was extracted from lipid biopsy samples using the RNeasy Lipid Tissue Mini Kit (cat. no. 74804), with a combination of a Qiagen TissueRuptor (Cat No. 9001273) and QIAShredder columns (Cat No. 79654) used for tissue homogenisation. After homogenisation each sample was split into two tubes, and a further 0.5ml of QIAzol reagent added to each tube. Each tube was then treated as a separate RNA extraction, following the RNeasy Lipid Tissue Mini Kit protocol as written. Once RNA was eluted the aliquots were recombined to give a final volume of 60ul eluted RNA. All RNA libraries were produced using the Beckman Coulter Biomek FxP (Cat No. A31842) with New England Biolab Ultra Directional RNA Library Prep Kit (Cat No. E7420L) and NEBnext Multiplex Oligos for Illumina Dual Index Primers (Cat No. E7600S), using provided protocols. 100ng of total RNA was used for library construction where possible. However in some cases the extracted RNA was at too low a concentration. For these samples 25ng total RNA was used for library construction. 13 cycles of PCR amplification were carried out upon all libraries of 100ng total RNA, and 15 cycles upon libraries with 25ng input total RNA.