92 Agilent 4150 TapeStation System Manual
7
Good Measurement Practices
Ladder Considerations
Ladder Considerations
The ScreenTape assays allow determination of the size of unknown sample
fragments or smear regions. This size determination is based on a ladder
which is a sample with specific sizing standards. The required ladder for an
assay is provided with the ScreenTape Reagents or can be purchased
standalone.
For best sizing precision and accuracy, a ladder per ScreenTape device is
recommended. Alternatively, an electronic ladder is also available for the
D1000 and High Sensitivity D1000, as well as the Cell-free DNA Screentape
assay. The electronic ladder can be selected in the TapeStation Controller
software and is added to the data automatically.
For the TapeStation systems, the ladder is exclusively loaded from location A1
on the tube strip holder. A ladder is typically prepared like a sample. For an
analysis that consists of runs with multiple ScreenTape devices a distinct
higher ladder volume is prepared. Please refer to the assay-specific Quick
Guide of the assay you plan to run for details on the preparation of this higher
ladder volume (Table 3 on page 64).
Some ScreenTape assays such as the standard D5000 and High Sensitivity
D5000 ScreenTape assay, as well as the Genomic DNA ScreenTape assay
require a ladder run per ScreenTape device. These assays do not offer usage of
electronic ladders. Running a ladder per ScreenTape device is a must in
analyses which involve multiple ScreenTape devices of these assays.
RNA ScreenTape assays require only one ladder per complete analysis even if
this analysis is involving multiple ScreenTape devices. For RNA assays an
electronic ladder can be selected in the TapeStation Controller software and
would then be added to the data automatically.
To select usage of an electronic ladder instead of a real ladder run on the ScreenTape
device, refer to inserting electronic ladder in “TapeStation Controller Software User
Interface” on page 53.
Typically, one lane of ladder is run for every ScreenTape device used. Higher volumes of
ladder are required for runs with multiple ScreenTape devices.
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