Prerequisite Protocols

• Routine Taq PCR

Materials

• PEG PCR Precipitation Buffer
• Ice Cold 80% Ethanol
• 1.5mL Centrifuge Tubes
• Sterile Distilled Water

Equipment

• Microcentrifuge
• Pipettes
• PCR Machine holding at 37C
• Static Incubator set to 37C
• DNA Quantification Device (optional but nice to have)

Step By Step

1. To a completed PCR reaction in a 0.2mL PCR tube, add 1:1 volume of PEG PCR Precipitation Buffer .
2. Vortex with 3 pulses of 1 second each to thoroughly mix.
3. Incubate at 37C in a PCR machine for 15 minutes.
4. Transfer to 1.5mL tubes and spin down 15mins at maximum speed (14,000rpm on my fuge). Align the hinges of the tubes so they face outward. This gives you a reference of where to look when searching for signs of pellet, or where to avoid sucking up to not disturb likely-invisible pellet.
5. Discard the supernatant carefully, ideally with a 200uL pipette tip with beveled tip.
6. Add 200uL of ice cold 80% Ethanol, vortex with 5 pulses of 1 second to mix, and spin down for 10mins at max speed (14,000rpm).