Agilent 4150 TapeStation System Manual 99
Good Measurement Practices
7
RNA Assays
RNA Assays
Heat Denaturation
The two RNA assays below include a heat denaturation step in the workflow.
• RNA ScreenTape assay
• High Sensitivity RNA ScreenTape assay
For heat denaturation, an accurately calibrated block heater, or a PCR
instrument fitting to 200 μL vials is required. This equipment is not supplied
with the TapeStation system. Before the heating step, close the tube strips or
96-well sample plates with lids or foil seal.
Heat denaturation typically consists of:
• Collecting sample and sample buffer at the bottom by centrifugation
• Incubation at 72 °C for 3 min
• Incubation on ice for 2 min
• Centrifuge to collect samples at the bottom of the vial
See also “Tools and Handling” on page 84 and “Troubleshooting RNA
Applications” on page 200.
Figure 25 Example of extra peaks being detected at the 28S position due to insufficient
denaturation in the Gel View (Left) and Electropherogram View (Right) of the
TapeStation Analysis software.
After each heating step, spinning down samples is required to collect condensed material
from the lid or foil seal.
Many heat blocks and PCR machines display a temperature that can be incorrect by up to
10 °C. Ensure that your heat block is appropriately calibrated before use.
To Next Page
Loading...
Need help?
General
