Leica Kreatech FISH probes Mode D'emploi page 11

Kreatech FISH probes
Minimum probe application:
Glass cover slip size
22 x 22 mm
22 x 32 mm
22 x 50 mm
Procedural recommendations:
Temperature and buffer concentration (stringency) of hybridization
and washing are important, as lower stringency can result in
non-specific binding of the probe to other sequences, and higher
stringency can result in a lack of signal. Incomplete denaturation of
target DNA and/or probe DNA can result in lack of signal.
Material required, but not supplied: Reagents:
1. Xylene
2. Formamide
3. Ethanol 100%, 85% and 70%
4. 0.01 M sodium citrate pH 6.0 or 8% sodium thiocyanate
5. 0.01 M HCl and 0.2M HCl
6. Pepsin Solution RtU (LK-101A)
7. 1 x PBS, pH 7.4
8. 2 x SSC, pH 7.0
9. Wash Buffer I (0.4 x SSC / 0.3% Igepal) (LK-102A)
10. Wash Buffer II (2 x SSC / 0.1% Igepal) (LK-103A)
11. DAPI counterstain (LK-095A (0.1 μg/ml) or LK-096A (1 μg/ml))
12. Counterstain diluent (LK-097A)
13. Fixogum (LK-071A) or rubber cement
Material required, but not supplied: equipment:
1. ThermoBrite (TS-01/02) or ThermoBrite Elite
(See www.LeicaBiosystems.com for more information)
2. Incubator at 37 °C
Minimum probe application
10 μl
15 μl
23 μl
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