Quick Protocol
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Additional protocol information in Technical Manual #TM673, available online at:
www.promega.com
Automated ccfDNA Purification
Preparing the Cartridge
1. Change gloves before handling Maxwell
®
RSC
Cartridges, RSC Plungers and Elution Tubes (0.5ml).
Place the cartridges to be used in the deck tray(s) with
well #1 (the largest well in the cartridge) facing away
from the elution tubes. Press down on the cartridge to
snap it into position. Carefully peel back the seal so that
all plastic comes off the top of the cartridge. Ensure
that all sealing tape and any residual adhesive are
removed before placing cartridges in the instrument.
2. Using a pipette, transfer 500µl of well #1 (the large
well) into the tube containing the magnetic resin pellet.
3. Resuspend the resin in the reduced volume. We
recommend using a pipette for resuspending the resin
because vortexing might cause resin to adhere to the upper
sides of the tube.
4. Transfer the resin and liquid back to well #1 of each cartridge (well #1 is the largest well).
5. Place one plunger into well #8 of each cartridge.
6. Place an empty 0.5ml Elution Tube into the elution tube position for each cartridge in the deck tray. Add 75µl of the appropriate
elution buffer to the bottom of each elution tube (see Notes below). This will give a final elution volume of approximately 60µl after
processing.
Notes:
a. We have developed two optimized elution buffer formulations for this kit. The NGS Elution Buffer (Cat.# MC1521) will elute
ccfDNA that is predominantly double-stranded. The PCR Elution Buffer (Cat.# MC1511) gives the most efficient elution that
works well in amplification-based assays. Ensure the selected elution buffer is at the bottom of the tube for optimal elution.
b. The NGS Elution Buffer is optimized for assays that require dsDNA. These assays include fluorescent dye quantitation,
electrophoresis and whole genome sequencing.
c. The PCR Elution Buffer is optimized for assays that use both ssDNA and dsDNA. These assays include quantitative PCR,
droplet digital PCR and amplicon-based sequencing.
7. Proceed to the next section, Running the Method on Maxwell
®
Instruments.
Running the Method on the Maxwell
®
Instruments (Cat.# AS4500, AS6000*, AS8500,
AS8000*)
1. To run the Maxwell RSC ccfDNA LV Plasma method, the appropriate Maxwell
®
RSC ccfDNA LV Plasma method must installed on
your instrument. Methods are available at:
www.promega.com/resources/software-firmware/
2. Follow the instrument run instructions in the
Maxwell
®
RSC ccfDNA LV Plasma Kit Technical Manual
#TM673.
*The Maxwell
®
RSC ccfDNA LV Plasma Kit is compatible with Maxwell
®
CSC Instruments operating in RUO mode.
Instructions for Use of Product AS1840.
Maxwell
®
RSC ccfDNA LV Plasma Kit
10828T
A
Figure 1. Setup and configuration of the deck tray.
The CSC/RSC plunger is placed in well
#8 of the cartridge (the well closest to the Elution Tube), and lysates and Lysis Buffer are
placed into well #1 of the cartridge.
