GSM3544636: plusDOX_G1_IL11RplusPLXDCplus; Mus musculus; RNA-Seq
Lungs of tumor bearing animals were dissociated, stained for IL11RA and PLXDC and sorted into lysis buffer (Qiagen RNeasy kit). RNA extraction was performed according to the manufacturer's protocol. cDNA libraries were constructed using Nextera XT DNA Sample preparation kit (Illumina) according to the manufacturer's recommendations with minor modifications. Briefly, cDNA amplicons were fragmented and barcoded by tagmentation using Tn5 DNA transposase. Tagmented cDNA were amplified with a limited-cycle PCR program. During the PCR step, indices 1 and 2 were added to allow for cluster generation and multiplexed sequencing. The extension time was increased from 30s to 60s. The amplified cDNA libraries were purified and cleaned using 0.6x Agencourt AMPure XP (Beckman Coulter) and fresh ethanol, and eluted in TE buffer. The quality of sequencing libraries was assessed using a high-sensitivity DNA chip (Agilent). Finally, the libraries were sequenced deeply using the HiSeq2000 sequencer (Illumina).
GEO Accession
GSM3544636