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Humidification of the Twin Towers for Microarray
or In Situ Hybridizations
The MJ Research Twin Towers slide cycler is well suited to multi-slide
hybridizations as well as slide thermal cycling. The highly accurate
and uniform temperature control (+/- 0.5°C) ensures very reproducible
results. Long-term hybridizations may be simply performed under
a glass cover slip if the slide chamber is maintained at high
humidity. The following simple procedure can maintain a humid
environment in the Twin Towers slide cycler for over 2 days.
Alternatively, hybridizations may be performed on slides in chambers
formed by adhesive frames and flexible polyester cover slips (MJ
Research Frame-Seal chambers). When using Frame-Seal chambers,
humidification of the Twin Towers is not necessary.
Preparing the humidification pad
- Obtain a sheet of thick blotting paper (e.g., VWR Grade 320
Blotting Paper) and cut pads 9.2 cm long by 2.5 cm wide.
- Holding one end, partially insert the pad into the lowest
slot in the slide tower.
- Add water to the pad to achieve saturation (about 3-4 ml).
- Insert the pad into the slot until it touches the rear panel,
then lightly bend down the pad in front until it touches the
base. When the door is closed, the pad will touch the clear
acrylic window. This collects and recirculates any condensate
that runs down the window.
Pre-heating and humidifying the Twin Towers
Usually a hybridization protocol calls for a denaturation step
prior to hybridization.
- For such a protocol, program the Twin Towers so that the first
step holds 95°C "forever". Follow this step with
a 95°C step for 2 minutes (or your desired denaturation
conditions), and then with the appropriate hybridization temperature
and time.
- While the slides are being readied, start the protocol and
allow the slide chamber to hold at 95°C until the chamber
is fully humidified (about 2-3 minutes).
- When the slides are ready, insert them into the Twin Towers,
close the door, and press "Proceed". This advances
the program to the second step and begins the hybridization
sequence according to the programmed conditions.
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