title: GSM4083677: T24-control-2; Homo sapiens; RNA-Seq
library_construction_protocol: total RNA was extracted using Trizol following the manufacturing protocol Library preparations were constructed according to the manufacturer's protocol. Briefly, The poly(A)mRNA isolation was performed using Poly(A) mRNA Magnetic Isolation Module or rRNA removal Kit.The mRNA fragmentation and priming was performed using First Strand Synthesis Reaction Buffer andRandom Primers. First strand cDNA was synthesized using ProtoScript II Reverse Transcriptase and the second-strand cDNA was synthesized using Second Strand Synthesis Enzyme Mix. The purified doublestranded cDNA by beads was then treated with End Prep Enzyme Mix to repair both ends and add a dAtailing in one reaction, followed by a T-A ligation to add adaptors to both ends. Size selection of Adaptorligated DNA was then performed using beads, and fragments of ~420 bp (with the approximate insert size of 300 bp) were recovered. Each sample was then amplified by PCR for 13 cycles using P5 and P7 primers, with both primers carrying sequences which can anneal with flow cell to perform bridge PCR and P7 primer carrying a six-base index allowing for multiplexing. The PCR products were cleaned up using beads, validated using an Qsep100 (Bioptic， Taiwan， China), and quantified by Qubit3.0 Fluorometer (Invitrogen, Carlsbad, CA, USA).Then libraries with different indices were multiplexed and loaded on an Illumina HiSeq instrument according to manufacturer's instructions (Illumina, San Diego, CA, USA).
attributes.tag: GEO Accession
attributes.value: GSM4083677