Washing; Washing In Hybridisation Bottles - Thermo Scientific Shake n Stack 6240 Operating And Maintenance Manual

Section 4
Methodology of Hybridisation

Washing

Washing in Hybridisation
Washing in a Sandwich
4-4 Shake 'n Stack
Two methods for washing follow.
1. Remove the bottle from the Oven.
Bottles
2. Pour off the hybridisation fluid and then half fill the bottle with the
first pre-warmed wash solution (approx. 3.38 fl. oz. [100 ml]) and
replace the cap.
Note Room temperature washing or washing with solutions which have
not been pre-warmed may result in background problems (see Appendix I
of the Hybridisation Guide at the end of this manual).
3. Replace the bottle in the Oven and leave for the required time period.
If transferring from sandwich box or plastic bag methodology, the
times and temperatures are usually those currently being used,
approximately 20 minutes per wash step.
4. Repeat Steps 1-3 for each additional wash. The wash solutions,
temperatures, etc., should be those recommended by the membrane
manufacturer, or refer to the Hybridisation Guide.
Note All wash solutions should be pre-warmed for best results.
Some scientists prefer to remove the membranes from the bottles and wash
them all in one container. Washing may be performed very simply in the
Box
Shake 'n Stack on the shaking platform and is particularly useful when
several membranes are being processed simultaneously.
1. Remove the hybridisation bottle from the oven. Pour off the
hybridisation fluid.
2. Using a pair of forceps, carefully remove the membranes and place
them in a suitable container with a lid (e.g. sandwich box or similar).
3. Using the forceps, gently unroll the membranes.
4. Add a sufficient volume of the pre-warmed first wash solution to
totally immerse the membranes. Replace the lid and shake on the
shaking platform in the Shake 'n Stack Oven at the required
temperature for the specified time period.
Note Initial room temperature washes are not recommended and may
result in subsequent background problems.
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