Leica Kreatech FISH probes Mode D'emploi page 15

Kreatech FISH probes
Probe preparation
Briefly spin down probe vial, vortex probe vial and briefly spin down
again before use. Allow probe to reach RT before use.
Research Use Only (RUO) Kreatech FISH probes are supplied Ready
to-Use (RtU).
Centromere, Subtelomere and Whole chromosome FISH probes are
provided in 5 x concentrated formats and must be diluted with the
provided hybridization buffer and/or other concentrated probes.
Consult label on vial and specific probe package insert for dilution
specifics.
Co-denaturation:
1. Apply 10 μl of probe or probe-mix per 22 x 22 mm field.
2. Cover with glass coverslip and seal with Fixogum (LK-071A) or
rubber cement.
3. Denature sample and probe on a ThermoBrite (TS-01/02) for 5 min
at 75 °C.
Hybridization:
Incubate overnight at 37 °C in a ThermoBrite (TS-01/02) or in a
humidified chamber.
Post-Hybridization Wash:
1. Pre-warm Wash Buffer I (0.4 x SSC / 0.3% Igepal) (LK-102A) to
72 °C.
2. Remove rubber cement.
3. Place up to 14 slides in 200 ml of Wash Buffer II (2 x SSC / 0.1%
Igepal) (LK-103A), incubate for 2 min at RT. Slide off coverslips.
Re-use only once for a total of 28 slides.
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