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Sequencing reaction cleanup
  1. 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
  2. Fill holes on Sephadex plate loader
  3. Invert Millipore multiscreen plate on plate loader. Invert plate and plate loader and tap plate loader to place Sephadex into wells on plate
  4. Add 300 μl dH20 to each well
  5. Let plate stand for 20-30min
  6. Centrifuge plate (balance with other plate or plate with only water) for 2min at 2500rpm
  7. Rotate plate 180º and spin for 2min at 2500rpm
  8. 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
  9. Add 15-20 μl dH20 to sequencing reaction
  10. Transfer sequencing reactions onto Sephadex columns on filter plate taking care not to damage columns
  11. Centrifuge plate for 2min at 2500rpm
  12. Rotate plate 180º and spin for 2min at 2500rpm
  13. Remove multiscreen filter plate, dump Sephadex into trash and rinse with dH20 for future re-use
  14. Dry sample plate in vacuum centrifuge at 60º for about 25-30min
  15. Cover plate with SealPlate
  16. Label appropriately, refrigerate and submit for sequencing

 
 

Kelly Miller Lab, Department of Biology, University of New Mexico, 167 Castetter Hall, MSC03 2020, Albuquerque, NM 87131-0001 USA
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