Promega AS1135 Maxwell RSC FFPE Plus DNA Kit Instruction Manual

Promega AS1135 Maxwell RSC FFPE Plus DNA Kit

Preprocessing of FFPE Section Samples

1. Place the FFPE tissue section into a 1.5ml microcentrifuge tube. If using slide-mounted tissue sections, scrape the section off the slide using a clean razor blade. Centrifuge the tube at maximum speed for 15 seconds to collect the sample at the bottom of the tube.
   Note: 5–10 micron-thick tissue sections ranging in size from 20mm2 to 200mm2 for a total of up to 2.0mm3 of tissue can be used.

2. Prepare Proteinase K by adding 500µl of Nuclease-Free Water to a tube of lyophilized Proteinase K (Part# V302A), for a final concentration of 20mg/ml Proteinase K. Dispense into aliquots and store at –20°C. Before use, thaw an aliquot and store on ice.

3. Add 20µl of 20mg/ml Proteinase K and 180µl of Incubation Buffer to a tube containing the tissue section.

4. Close the tube cap and heat the sample at 70°C for 1 hour overnight.
   Note: Samples incubated overnight are likely to yield more amplifiable gDNA.

5. Add 400µl of Lysis Buffer to each sample.

6. Vortex samples and centrifuge briefly to collect lysate at the bottom of the tube.

7. Transfer lysate to well #1 (the largest well) of the Maxwell® RSC Cartridge or save until ready for automated DNA extraction using a Maxwell® Instrument.

Maxwell® Automated DNA Purification

Cartridge Preparation

1. Place the cartridges to be used in the deck tray(s) with well #1 (the largest well) facing away from the elution tube.

2. Press down on the cartridge to snap it into position. Ensure both cartridge ends are fully seated in the deck tray. Carefully peel back the seal so that the entire seal is removed from the top of the cartridge. Ensure that all sealing tape and any residual adhesive are removed before placing cartridges in the instrument.
   Caution: Handle cartridges with care. Seal edges may be sharp.

3. Place a plunger in well #8 of each cartridge. Well, #8 is the well closest to the elution tube.

4. Place an empty elution tube into the Elution Tube position for each cartridge in the deck tray(s).

5. Add 50μl of Nuclease-Free Water to the bottom of each Elution Tube. The elution tubes must stay open during the DNA purification.
   Notes: Use only the Elution Tubes and Nuclease-Free Water supplied with the Maxwell® RSC FFPE Plus DNA Kit. The use of other elution buffers may impact DNA purification performance or downstream use.

Instrument Run on the Maxwell® Instruments

Follow the instrument run instructions in the Maxwell ® RSC FFPE Plus DNA Kit Technical Manual #TM574.

Maxwell is a registered trademark of Promega Corporation.

Additional protocol information is in Technical Manual #TM574, available online at: www.promega.com

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