I'm getting poor quality/poor purity of RNA after isolation with the RiboPure RNA Purification kit. What could be the cause of this?
Poor quality/poor purity RNA is typically due to the following:
- Partially degraded RNA: Use fresh tissue or cells, or use RNAlater Stabilization Solution if tissue cannot be frozen immediately.
- DNA contamination: If the sample contains organic solvents, strong buffers, or an alkaline solution, we recommend performing phase separation at 8 degrees C.
- A low A260/A280 ratio (<1.6): This indicates that an insufficient amount of the TRI Reagent was used. Incubation of the homogenate at room temperature for 5 minutes can help nucleoproteins dissociate from RNA. Phenol contamination could also cause a low absorbance ratio.
What is the column cut-off size for the RiboPure RNA Purification Kits?
The column cut-off size is 200 nt. Using a RiboPure kit is not appropriate for miRNA recovery. For miRNA isolation, we would suggest using a mirVana kit, the TRIzol Plus RNA Purification Kit, or the PureLink RNA Micro Kit.