UM_TotalRNA_NucleoZOL(18)
时间:2025-07-11
时间:2025-07-11
user's manuals about RNA or or
6.2 Troubleshooting
Problem
Possible cause and suggestions
Homogenization or sample lysis is incomplete
Insufficient yield
Improve homogenization by testing more stringent conditions.
Increase volume of RNase-free water for dissolving of RNA and prolong mixing for solubilization. Increase volume of NucleoZOL.
Use Buffer AE (5 mM Tris pH 8.5) for sample dilution for spectrophotometric quantification.
Increase volume of RNase-free water for dissolving of RNA and prolong mixing for solubilization.
Perform phase separation as described in section 5.2.3.Make sure to use fresh tissue or flash-freeze tissue immediately upon harvest.Store samples at -70 °C.
Make sure cells stay intact during trypsinization.Make sure to work in an RNase-free environment.
Solubilization of the RNA pellet is incomplete.
Volume of NucleoZOL used for homogenization was too low
Low pH during spectrophotometric quantification
Ratio A260/280 < 1.6
RNA pellet was only partly solubilized
Contamination of polysaccharide or proteoglycan
Inadequate tissue sampling
Inappropriate storage conditions
Degraded RNA
Cell dissolution during trypsinizationRNase contaminated solutions or tubes