DV200 Analysis of FFPE RNA Samples with the 2100 Bioanalyzer System

Answer

This Information Applies To: 2100 Bioanalyzer, 2100 Expert software

Issue

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. The DV₂₀₀ of FFPE RNA samples can be calculated within the Agilent 2100 Expert software.

Steps to follow

Software version B.02.10 or higher

If you are using 2100 Expert software version B.02.10 or higher, the DV₂₀₀ RNA Nano and Pico assays will be available to run from the Assay Selection drop-down list (Figure 1). The software version can be displayed in the 2100 Expert software by clicking Help>About 2100 Expert.... If you are using an older version, you can run the DV₂₀₀ assays by upgrading to the latest software version. The latest software is available to download free of charge here and this article explains how to install it. The DV₂₀₀ RNA Nano and Pico assays will then be available in the software. An example DV₂₀₀ region in the 2100 Expert software is shown in Figure 2.

Figure 1. Selecting the DV₂₀₀ assays from the Assay Selection drop down menu

Figure 2. An example DV₂₀₀ region taken from the Demo DV₂₀₀RNA Nano file

Software version lower than B.02.10

If you are using an earlier software version and are unable to upgrade to the latest software version, download the DV₂₀₀ assay files:

Extract and store the DV₂₀₀ RNA Nano.xsy or DV₂₀₀ RNA Pico.xsy file in the 2100 Expert Software RNA assay folder: C:\Program Files (x86)\Agilent\2100 bioanalyzer\2100 expert\assays\RNA. The RNA Nano or DV₂₀₀ RNA Pico assays will then be available in the Assay Selection drop-down list (Figure 2).

Applying the DV₂₀₀ calculation to a previously run data file

If you have 2100 Expert software version B.02.10 or higher installed or have saved the DV₂₀₀ assay files, the DV₂₀₀ calculation can be applied to a previously run data file:

Open the data file that you want to apply the DV₂₀₀ to with the 2100 Expert software.
Navigate to the Assay Properties tab.
Click Import Setpoints. A dialog box will open.
Select the DV₂₀₀ RNA Nano or DV₂₀₀ RNA Pico .xsy file, depending on the data file open (Figure 3). The DV₂₀₀assay parameters will be applied to the data file.