title: GSM1194691: Patient 3 tumor 2; Homo sapiens; RNA-Seq
library_construction_protocol: Tissues were sectioned at a 7-micron thickness and mounted on regular uncharged glass slides for patients 1, 2, and 3, and on polyethylene napthalate (PEN) membrane slides (Leica) for patient 4, followed by H&E staining. LCM was performed using the Arcturus eIIx for patient 1 and 2, Zeiss PALM for patient 3, and Leica LMD7000 for patient 4. A tissue area of 800,000 to 1,200,000 μm2 was dissected and collected from each lesion.  RNA was extracted from laser-microdissected cells using the RNeasy Micro Kit (QIAGEN). The cDNA was generated using the Ovation RNA-Seq System (NuGEN) for patients 1 and 2 and the 15 Ovation RNA-Seq V2 System (NuGEN) for patients 3 and 4. For patients 1 and 2, cDNA of ~200 bp was selected by gel purification. For patients 3 and 4, the cDNA was sheared to 140-180 bp using the Covaris focused-ultrasonicator with the following settings: duty cycle 10%; intensity 5; cycles per burst 200; total time 6 minutes.
attributes.tag: GEO Accession
attributes.value: GSM1194691