Large Scale mRNA Purification
mRNA from large scale extractions was purified by oligo-dT enrichment
utilising an ÄKTA pcc chromatography system (Cytiva). 10ml total RNA was
bound to a 1 ml volume Oligo-dt(18) column (Sartorius) in binding buffer
(50 mM Sodium Phosphate pH 7, 500 mM NaCl). The column was washed in 20
column volumes of wash buffer (50 mM Sodium Phosphate, 150 mM NaCl).
Bound mRNA was eluted in 5ml of water, before quantification by Nanodrop
spectrophotometer.