Leica Kreatech FISH probes Mode D'emploi page 18

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working order.
I have high background or a-specific cross-hybridization when using
Centromeric or Subtelomeric probes.
• Increase the temperature of the stringency wash (Wash Buffer I)
when washing slides.
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. FISH reagent Kit (KI-60005)
2. FISH digestion Kit (KI-60006)
3. 1% buffered formaldehyde / 1 x PBS / 20 mM MgCl
4. 1 x PBS, pH 7.4
5. 2 x SSC, pH 7.0
6. Pepsin solution (LK-101A)
7. Wash buffer I (0.4 x SSC / 0.3% Igepal) (LK-102A)
8. Wash buffer II (2 x SSC / 0.1% Igepal) (LK-103A)
9. Igepal
10. 0.01 M HCl
11. Carnoys fixative (methanol : acetic acid = 3 : 1)
Minimum probe application
10 μl
15 μl
23 μl
Kreatech FISH probes
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