title: GSM4043540: GInt1APC; Mus musculus; RNA-Seq
library_construction_protocol: 1. Sort into clear RNase free tube containing 200ul of Trizol, use 27G needle to homogenize cells. 2. Filled samples up to 1ml with Trizol, and added 200µl chloroform per 1ml of Trizol to each sample. 3. Incubation 10 min and Centrafuge. 4.Transferred 400µl of the upper aqueous layer to a new tube repeat step 2-3.  5.Transferred ~400 of upper aqueous layer to the tube had 1.5µl glycogen. 6. Added 450µl Isopropanol to each sample and mix well. 7. Incubation 10 min and Centrafuge.  8.  Removed supernatant and Added 1ml 75% EtOH/DEPC H2O. 9. After centrafuge, removed supernatant and circled pellet with marker, air dried for 20 min. 10. Add adequate amount f DEPC H2O to each sample for 10min, mix well and keep at-80 ℃ until use. Libraries were prepared according to manufacturer's instructions using the TruSeq Stranded mRNA Prep Kit (Illumina) and NEXTflex DNA barcodes (Perkin Elmer). The resulting libraries were purified using the Agencourt AMPure XP system (Beckman Coulter) then quantified using a Bioanalyzer (Agilent Technologies) and a Qubit fluorometer (Life Technologies).  Libraries were re-quantified, normalized, pooled and sequenced on an Illumina NextSeq 500 instrument as High Output 75bp single read runs. Following sequencing, Illumina NextSeq Real Time Analysis version 2.4.11 and bcl2fastq2 version 2.20 were run to demultiplex reads and generate FASTQ files.
attributes.tag: GEO Accession
attributes.value: GSM4043540