How to Determine the DV200 of FFPE RNA Samples on the Agilent TapeStation

Created by Geoff Pettitt 6 months ago | Last modified by Geoff Pettitt 3 months ago

This Information Applies To: 4200, 4150 and 2200 TapeStation, TapeStation analysis software A02.02 or higher.

Issue:

When using the Agilent 4200, 4150 and 2200 TapeStation systems, the DV₂₀₀ of FFPE RNA samples can be calculated within the TapeStation analysis software. The DV₂₀₀ score is a quality score for evaluating quality of RNA derived from formalin-fixed paraffin-embedded (FFPE) samples established by Illumina Inc. in 2016. It represents the percentage of RNA fragments larger than 200 nucleotides relative to all RNA fragments.

Steps to follow

The workflow to calculate the DV₂₀₀ described here applies to software version A.02.02 or higher. If you are using an earlier version, download the 2200 TapeStation software or 4200/4150 TapeStation software. To apply the DV₂₀₀ calculation to an existing data file (Figure 1):

Open the data file that you wish to apply the DV₂₀₀ metric to and click Home>Region.
Click Region Settings.
Enter the lower (e.g. 200 nt) and upper limits (e.g. 10,000 nt) of the region in the dialog box. You may type DV₂₀₀ as region comment so that the region can be identified in the region table later.
To apply the DV₂₀₀ region to all samples in the active file, select This File. To save a DV₂₀₀ region on this PC that will be applied to all files of this assay type that are opened on this PC, select Assay Files.
Click OK or Apply.