The beads were isolated using a magnet, washed, mounted on a glass slide and imaged by confocal microscopy.
Cells attached to beads were isolated using a magnetic particle concentrator (Dynal, Merseyside, UK) and cultured for 6 hours at 37°C.
PolyA+ RNA was isolated using beads from TruSeq RNA Sample Prep Kit v2 (Illumina kit RS-122-2001), and PolyA+ RNA-seq libraries were prepared using Epicentre ScriptSeq v2.
Snap frozen, fresh tissue was homogenized using Lysing Matrix A (MP Biomedicals) and a mini bead-beater (Biospec Products) and RNA was isolated using the RNeasy kit (Qiagen).
Total RNA was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, US) and treated with RNase-free DNase I. Poly(A) mRNA was isolated using oligodT beads and fragmented into small pieces in Thermomixer under elevated temperature.
Cells were homogenized and lysed in lysis/binding buffer and mRNA was isolated using magnetic beads (oligo (dT 25) as described by the manufacturer (Dynal AS, Oslo, Norway).
Total RNA from frozen samples was isolated using zirconia/silica beads (0.5 mm diameter; Biospec, Bartlesville, OK, USA) and a Mini-Beadbeater-96 (Biospec) with 1 mL TRIzol reagent (Invitrogen/Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions.
Total RNA from frozen samples was isolated using Zirconia/Silica beads (0.5 mm diameter; Biospec) and a MiniBeadbeater (Biospec, Bartlesville, USA) with 1 mL TRIzol reagent (Invitrogen Life Technologies, Carlsbad, CA, USA) according to the manufacturer's protocol.
Total RNA from frozen samples was isolated using Zirconia/Silica beads (0.5 mm diameter; Biospec Products, Bartlesville, USA) and a Mini-Beadbeater (Biospec Products, Bartlesville, USA) with 1 mL TRIzol reagent (Invitrogen, Grand Island, USA) according to the manufacturer's protocol.
DNA was isolated using a beat-bead procedure using 2-4 glass beads (2.5 mm) to accomplish cell lysis and subsequent DNA purification using the FastDNA® SPIN Kit (MP Biomedicals, Solon, OH).
Next, Poly (A) mRNA was isolated using oligo-dT beads (Qiagen).
