Zymo cleanup and concentration

Zymo Genomic DNA Clean & Concentrator - 10 Protocol

1. Add 5 volumes of ChIP DNA Binding Buffer to each volume of DNA sample. Mix thoroughly and spin down.

   e.g., to 100uL of sample DNA, add 500uL ChIP DNA Binding Buffer

2. Transfer mixture to a Zymo Spin Column (IC-XC) in a collection tube.

3. Centrifuge 30s at 12,000 RPM. Discard flow through.

4. Add 200uL DNA Wash Buffer to the column. Centrifuge 1 min at 12,000 RPM. Discard flow through.

5. Repeat Step 4.

6. Add >= 10 uL DNA Elution Buffer (or nuclease-free water) directly to the column matrix and incubate at room temp for 1 minute. Transfer column to a labelled 1.5mL tube and centrifuge for >=30 sec to elute DNA.

   To improve yield, one can heat DNA Elution Buffer to 60C before use.

Notes:

• Aliquoted DNA Elution Buffer to 2mL tube and heated to 60C before use.

• Eluted each sample in 50uL of buffer.

Performed Zymo Cleanup on both Round 1 and Round 2 extractions at 1:00pm, July 12, 2025. Stored cleaned DNA in fridge for the weekend.