title: GSM3099012: timeseries sample441; Homo sapiens; OTHER
library_construction_protocol: Cells were collected by centrifugation (300xg, 5 min) and the genomic DNA was isolated using the Isolate II Genomic DNA Kit (Bioline). PCR was performed in two steps; PCR1 with ~100 ng genomic DNA and site specific barcoded primers. PCR2 used 2 µL of each PCR1 product with Illumina PCR Index Primers Sequences 1–12. Each sample was generated with a unique combination of a barcode and index. Both PCR reactions were carried out with 25 µL MyTaq Red mix (Bioline), 4 µM of each primer and 50 µL final volume in a 96 well plate. PCR conditions were 1 min at 95 ˚C, followed by 15 sec at 95 ˚C, 15 sec at 58 ˚C and 1 min at 72 ˚C (15x). 20 µL of 8 samples were pooled and 100 µL was loaded onto a 1% agarose gel. PCR product was cut from gel to remove the primer dimers and cleaned with PCR Isolate II PCR and Gel Kit (Bioline).
attributes.tag: GEO Accession
attributes.value: GSM3099012