RiboPure™ RNA Purification Kit, yeast

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Samples can be flash frozen (-80C) before use, this will likely cause lower RNA yield.		The isolated RNA was treated with RNAse-free DNAse I. When checking the yield by means of a spectrophotometer, use the supplied TE buffer to zero the machine

Upstream tips
Samples can be flash frozen (-80C) before use, this will likely cause lower RNA yield.
Downstream tips
The isolated RNA was treated with RNAse-free DNAse I. When checking the yield by means of a spectrophotometer, use the supplied TE buffer to zero the machine

Publication protocol

Total RNA isolation was done by using the components and the protocol of the RiboPure Yeast RNA Purification Kit (Life technologies). This included cell disruption with RNAse-free zirconium beads in the presence of RNAse inhibiting lysis solution and column purification. The isolated RNA was treated with RNAse-free DNAse I. To insure that another mode of isolation gave us similar results as the kit, RNA was extracted simultaneously once, using the hot acid phenol isolation protocol [11]. RNA quantification was done on total freshly isolated RNA, always diluted in the same volume of 50 μL, using a Qubit 2.0 fluorometer. To see if any polyadenylated rRNAs are involved, we did a one time poly-A selection at times T0 T30 T60 utilizing the BcMag Quick mRNA Purification Kit (Bioclone Inc.) according to the manufacturer’s directions.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for RiboPure™ RNA Purification Kit, yeast below.

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