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Sequencing reaction cleanup
- Obtain 96 well Millipore multiscreen filter plate. Add a used v-bottom 96 well storage plate to the bottom of filter plate using blue centrifuge alignment frame
- Fill holes on Sephadex plate loader
- Invert Millipore multiscreen plate on plate loader. Invert plate and plate loader and tap plate loader to place Sephadex into wells on plate
- Add 300 μl dH20 to each well
- Let plate stand for 20-30min
- Centrifuge plate (balance with other plate or plate with only water) for 2min at 2500rpm
- Rotate plate 180º and spin for 2min at 2500rpm
- Remove used 96 well storage plate and replaced with a new v-bottom 96 well storage plate taking care to ensure that rows and columns are aligned with Millipore multiscreen filter plate
- Add 15-20 μl dH20 to sequencing reaction
- Transfer sequencing reactions onto Sephadex columns on filter plate taking care not to damage columns
- Centrifuge plate for 2min at 2500rpm
- Rotate plate 180º and spin for 2min at 2500rpm
- Remove multiscreen filter plate, dump Sephadex into trash and rinse with dH20 for future re-use
- Dry sample plate in vacuum centrifuge at 60º for about 25-30min
- Cover plate with SealPlate
- Label appropriately, refrigerate and submit for sequencing
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Kelly B. Miller Lab, Department of Biology, University of New Mexico, 167 Castetter Hall, MSC03 2020, Albuquerque, NM 87131-0001 USA
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