GSM3873025
GSM3873025: Myc_NeuNT_MGT_14; Mus musculus; RNA-Seq
Total RNA was extracted with Trizol according to manufacturer's protocol. Sequencing libraries were constructed using an Illumina TruSeq RNAseq kit. Briefly, poly(A)+ RNA was isolated from 1ug of total RNA (per sample) using oligo-dT coated magnetic beads. The recovered RNA was then chemically fragmented. First strand cDNA was generated using random hexamers as primers for reverse transcriptase. Following second strand synthesis, the ends of the double stranded fragments were repaired and then a single “A” nucleotide was added to each end. Illumina adaptors were ligated to the cDNAs. Limited round (11-cycle) PCR was used to amplify the material to yield the final libraries. Library concentration was determined using real time PCR with primers complementary to the Illumina adaptors. Sample libraries were diluted and applied to an Illumina single read flow cell at a concentration appropriate to generate about 180 million reads per lane. All  libraries were prepared with indexing barcodes to permit multiplexing in a single lane. 100 cycle single read sequencing was used to generate base call files. Illumina's CASAVA package was used to assemble the reads into standard fastq formatted data.
GEO
GSM3873025
GEO Accession
GSM3873025