|
Purpose: The original Brown lab protocol lists these steps
as optional. We do them immediately after the DNAs are spotted onto
the glass slides. The purpose of rehydration is to make the spots
more uniform and the purpose of cross-linking is to help bind the
DNA to the glass surface. The third treatment of blocking can be
done at a later time. Always avoid dust in these steps.
- Print ID and slide ID numbers: _______________________
Name and date: __________________________________
See the Protocol: Microarray Construction and Design for the
details of printing.
Ideally, treatments I (Hydration) and II (cross-linking) should
be done just after the printing is finished. On a very humid day,
you may have to first snap dry the spots on a heating block at
the end of the printing and before you proceed with the rehydration
(see the microarray construction protocol for details).
Post Print slide treatment I: Rehydrate and dry the dried spots
to make them more uniform:
____1. Heat water in a microwavable plastic beaker. Pass dry slides,
array side down, one-by-one the over hot water bath (70-90c) just
until you see fine mist (no big droplets) over arrayed area.
____2. Place moistened slide on flat heating block at 80-100c
(setting #4) for few seconds until moisture is just driven off.
____3. Set aside to cool on a clean tray. Keep covered with foil.
Post Print slide treatment II: UV cross-link the DNAs to the slide
coating:
____1. Place dry slides, grid side up on a paper towel
in StrataLinker
____2. Turn on power.
____3. Press 'Auto Cross Link'
____4. Set to 650 (=65mJ).
____5. Press 'Start'
____6. Beeps when finished. Press 'Reset' and turn power off.
____7. Store the slides in a dust free box labeled "Ready to Block"
and also labeled with the Print ID and Slide ID numbers
Post Print slide treatment III: Blocking of amine or aldehyde
slides
Follow the detailed protocols as needed for each type of slide:
Blocking cDNA arrays printed on amine slides (poly-L-lysine
or silanated) or Blocking cDNA arrays printed on aldehyde slides
(silylated)
|
