11 In Situ Hybridization for RNA: Nonradioactive Probe: Oligo-DNA Probe
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Results
Dot blot hybridization with T-T dimerized probes
Dot blot hybridization using T -T dimerized sense and antisense
KGFR oligo-DNA probes was shown in Fig 1. Aliquots (2/l1) of
KGFR sense oligo-DNA solution were dotted on nitrocellulose
filters at 1 pg to 10 ng /spot. When T -T dimerized KGFR antisense
oligo-DNA probe was hybridized with KGFR sense oligo-DNA
on a nitrocellulose filter, at least 10 pg of the oligo-DNA was detected, while no signal was seen with dotted KGFR sense oligoDNA.
In situ hybridization with T-T dimerized probes
In situ hybridization using T -T dimerized Metallothionein oligoDNA probes and 28S rRNA complementary oligo-DNA probe in
paraffin sections of esophageal cancer are shown in Figure 2.
10
100 10
ng ng
pg
pg pg
KGFR - 1 anti sense
KGFR 1 sense
Fig. 1. Colorimetric dot blot hybridization. Various amouts (1 pg to lOng/spot)
of KGFR sense oligo-DNA, which were fixed onto nitrocellulose membranes,
were hybridized with T-T labeled KGFR antisense oligo-DNA or T-T labeled
KGFRsense oligo-DNA. At least 10 pg of the antisense oligo-DNA was detected.