Preparation of FFPE DNA and RNA Samples for Sequencing Using an Automated Purification Instrument (API)

To begin, cut the formalin-fixed paraffin-embedded or FFPE DNA and RNA samples. Turn on the automated purification or API instrument and sign in with the username and password. Next, in the Integrated Genomic Viewer software, under samples, click Create Samples"enter the sample name and select the application category.

After entering the sample details, click on Save"Then select Runs"click on Sample to Result"and enter the run name before clicking Next"Select the assay followed by a sample of interest and click Next"After selecting each sample, click Assign"then check the elution parameters and save the run. Place FFPE tissue samples and sample processing tubes. Centrifuge the tube at 2, 000 G for one minute.

Add the Protease Digest master mix prepared from a nucleic acid purification kit into the tube. Incubate the sample at 60 degrees Celsius for one hour, followed by incubating it at 90 degrees Celsius for one hour. After cooling the samples, lift the inner tube of the sample processing tube and lock it by turning left.

Centrifuge the inner tube at 2, 000 G for 10 minutes. Then unlock the tube by turning right and separating it from the other tube. Discard the inner tube and place the outer tube on ice until loading on the API.

Prepare a DNA digestion master mix and load in prefilled FFPE DNA and RNA Purification plate two. Load the prepared samples in FFPE DNA and RNA purification plate one. On the API screen click Run"then select the run plan and click Next"Follow the on-screen instructions to load the required consumables and reagents for the purification run.

Once all the reagents are loaded, click Next"close the API door and click Start"T the end of the run, click Unload"and immediately remove the 48-well nucleic acid archive plate containing the purified DNA and RNA. After the sample purification is over, seal the plate and store it at minus 80 degrees Celsius.