CAMbank: cfDNA BCT Field Processing v1

After venipuncture, invert the tubes gently 8 to 10 times to fully mix tube anticoagulant with blood sample.

Store the tube upright at room temperature until centrifugation.

Note: The tube stabilizes cell-free DNA for up to 14 days at 6 °C to 37 °C and CTCs for up to 7 days at 15 °C to 30 °C.

Note: A swing bucket centrifuge is required.

Set centrifuge:

• acceleration: 9
• deceleration: 9
• temperature: RT
• duration: 20 minutes
• speed: 300xg

Place the cfDNA BCTs in the centrifuge.

Place a protective cover over the swing buckets in case of tube leakage.

Start the centrifuge.

Stand by the centrifuge until the centrifuge reaches max speed. Listen for signs of imbalance or compromised tube integrity.

Carefully remove cfDNA BCTs from the centrifuge and inspect for separation of red blood vs plasma layers.

Transfer cfDNA BCTs to a sanitized laminar flow hood.

Remove the upper plasma layer and transfer to a new 15mL conical tube.

Cap the conical tubes and return to the centrifuge.

Note: A swing bucket centrifuge is required.

Set centrifuge:

• acceleration: 9
• deceleration: 9
• temperature: RT
• duration: 10 minutes
• speed: 5000xg

Place the conical tubes in the centrifuge.

Place a protective cover over the swing buckets in case of tube leakage.

Start the centrifuge.

Stand by the centrifuge until the centrifuge reaches max speed. Listen for signs of imbalance or compromised tube integrity.

Recap and freeze the cfDNA tubes to preserve DNA in the red blood cell pellet

Carefully remove conical tubes from centrifuge.

Transfer conical tubes to a sanitized laminar flow hood.

Using a P1000, carefully aliquot plasma into 2D barcoded tubes at 500µL each.

Return 2D tubes with plasma to the rack and place in the -80C freezer.