vProbe labeling:

    Several methods are used to do probe labeling: Nick translation, Random Priming, and PCR.

Probes Labeling for FISH by Nick Translation

Materials:

Nick Translation Kit (Gibco, Cat#: 18247-015)

Components:

10X dNTP Mix

0.2 mM         each dCTP, dGTP, dTTP
0.1 mM         dATP
0.1 mM         biotin-14-dATP
500 mM        Tris-HCl (pH 7.8)
50  mM         MgCl2
100 mM        beta-mercaptoethanol
100 ug/ml      nuclease-free BSA

10X Enzyme Mix

0.5 U/μl         DNA Polymerase I
0.007 U/ul     DNase I
50 mM          Tris-HCl (pH 7.5)
5 mM            magnesium chloride
0.1 mM         phenylmethylsulfonyl fluoride
50% (v/v)      glycerol
100 ug/ml      nuclease-free BSA

Stop Buffer 0.5 M EDTA (pH 8.0)

Distilled H2O

Procedures:

 1.  Place a 0.6mL microcentrifuge tube on ice and allow the tube to cool.
2.  Pipet the following components to the tube: