Add ethanol (96-100 % ) to DNA Wash Buffer before use (120 mL ethanol to the DNA Wash buffer concentrate for a total volume of 150 ml). Note on the label that ethanol has been added.
Note all centrifugation steps are carried out at 3,000xg at RT.
- Add 5 volumes DNA Binding Buffer to 1 volume of sample and mix.
DNA Binding Buffer contains an integrated pH indicator. DNA adsorption requires a pH of ≤7.5, and the pH indicator in the buffers will appear yellow in this range. If the pH is >7.5 the binding mixture will turn orange or violet meaning the pH of the sample exceeds the buffering capacity of the DNA Binding Buffer and DNA adsorption will be inefficient. In these cases, add 10 μL 3M sodium acetate, pH 5.2, to adjust the pH of the binding mixture. The colour of the mixture should turn yellow.
- Place a Chromatrap® 96 DNA Purification plate and position on to the 96-well collection plate provided.
- Transfer each sample to a corresponding well on the Chromatrap® 96 DNA Purification plate.
- Centrifuge at 3,000xg for 60 seconds at RT. Discard the flow through.
- Add 700 μL DNA Wash Buffer to each well and centrifuge at 3,000xg for 60 seconds at RT. Discard the flow through.
- Spin dry at 3,000xg for 30 seconds at RT to remove any remaining liquid from the plate.
- Transfer the Chromatrap® DNA Purification plate on to a clean supplied 96-well DNA Elution plate.
- To elute DNA, add 50 μL DNA Elution buffer to the centre of the membrane of each well and incubate for 1 minute. Centrifuge at 3,000xg for 60 seconds to collect the eluted DNA.
Troubleshooting Guide and FAQs are given in the delivered protocol.