title: GSM2723940: BPS2; Homo sapiens; OTHER
library_construction_protocol: Total RNA was extracted using Trizol reagent. 50 μg total RNA was mixed with 250 pmol of the W2 probe (5’-CTTCTTAGGAGCTGTCCGAG-3’). RNA and probe were heated to 95°C for 5 min and then allowed to slowly cool to room temperature. 20 μl MyOne Streptavidin C1 Dynabeads (ThermoFisher Scientific) beads washed and resuspend in 2x streptavidin binding buffer as described above. RNA annealed to probe and beads were mixed and rotated at room temperature for 30 min. Beads were washed with wash buffer as described in section 3.5.8. Isolated transcripts were released from the beads by resuspending the beads in 100 μl wash buffer, heating to 95°C for 5 min and immediately adding to a magnet and collecting the supernatant. Isolated transcripts were then reprecipitated and resuspended in 20 μl water. Transcripts were then subject to an Iso-seq cDNA library preparation using the SMARTer PCR cDNA synthesis kit (Clontech) and size selected for 800 - 2 kb and 2 kb+ sized transcripts. SMRTbell adapters were then added to the isolated libraries.
attributes.tag: GEO Accession
attributes.value: GSM2723940