互联网2013-11-13
1. Protocol for total RNA purification from tissue sample with E.Z.N.A.® Total RNA System
1) Remove sample from RNASafer® Reagent using forceps.
2) Estimate the amount of the tissue and select appropriate E.Z.N.A.® Total RNA Kit. Use RNA Miniprep Kit (R6634) for less than 40mg tissue, Total RNA Midiprep kit (R6664) for 200mg tissue, RNA Maxiprep Kit(R6693) for 1 gram tissue.
3) For RNA isolation using RNA miniprep kit, add 700ul TRK lysis buffer. Homogenize the tissue with a rotor-stator homogenizer. For RNA isolation using Midi and Maxiprep kit, follow the standard protocol in the user manual.
4) Follow standard protocol for E.Z.N.A.® Total RNA Kits in user manuals.
2. Protocol for total RNA purification from culture cells and white blood cells with E.Z.N.A.® Total RNA System
1) For RNA isolation using RNA miniprep kit, take 150ul cell/RNASafer® Reagent mixture to a RNase free 1.5 ml tube. Add 200ul TRK lysis buffer and mix with 350ul 70% ethanol. For RNA isolation using Midi and Maxiprep kit, follow the standard protocol in the user manual.
2) Apply the sample to the HiBind® RNA column and follow standard protocol in user manual for each kits.
When working with RNASafer® Reagent use gloves and eye protection (safety goggles) and avoid contact with skin or clothing. Unless otherwise noted, all steps are to be carried out at room temperature (20°C -25°C).
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