Team:Freiburg/Protocols/PDMS PDITC

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PDMS-PDITC

protocol for PDITC surface on flow cells
According to protocol from AG Roth

material: chemicals, used kits, …
duration: … min

Slide preparation

  1. Name the slide
  2. Clean slides by washing with ethanol and rubbing (per hand, with glove)
  3. flush with aqua dest.
  4. Dry with wafergun
  5. Activate in plasma generator: gas flow 80 L/h, time 1 min

APTES

  1. Prepare APTES-solution: for 5 slides:
    • 18 mL ethanol (90%)
    • 1 mL dd aqua (5%)
    • 1 mL APTES (5%)
  2. Incubate slides over night at RT in slideholder (transparent, sealed with parafilm) filled with APTES-solution
  3. Wash each flow cell individually with ethanol
  4. immerse for 5 min in ethanol in slideholder
  5. quickly dispose ethanol and dry flow cells with wafergun
  6. Put slides in new slideholder and bake them in oven at 70 °C for 60-90 min

PDITC

  1. Prepare PDITC-solution: for 5 slides:
    • pyridin 2.0 mL
    • DMF 18.0 mL
    • PDITC 40 mg
  2. Cool down slides in slidebox with wafergun (go close with gun)
  3. Incubate slides in slideholder for 2 h filled with PDITC-solution at RT
  4. Discard PDITC-solution
  5. Fill with ethanol and discard ethanol
  6. Wash 2 x 5 min in slideholder with ethanol
  7. Fill with ethanol and discard ethanol once more
  8. Dry with wafergun
  9. stock in N2 atmosphere in box wrapped with parafilm
  10. storage >5 days
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