Chapter 11
In Situ Hybridization for RNA: Nonradioactive
Probe: Oligo-DNA Probe: T -T Dimer
YOSHITAKA HISHIKAWA, MASAMOTO MURAKAMI,
MASAO KASAHARA AND NAOFUMI NAGASUE
Introduction
The method using thymine-thymine (T-T) dimerized DNA as a
haptenic probe was previously reported (Koji 1988, 1989, 1990,
1996) and we have elaborated it in the following sections.
When ultraviolet rays (UV: 254nm) are exposed to the location adjacent to thymidines in the DNA strand, Pyrimidine ring
of thymidine is detached and then new skeletal structure between both thymidines is constructed. This is called T -T dimer,
which is very stable and has antigenic property. Using this method takes advantage of the fact that T -T dimer formation in DNA
is proportional to the dose ofUV irradiation (Franklin 1982) and
the frequency of occurrence of two successive thymine bases in
the DNA, and that no purification step for isolating T -T dimerized DNA is required. Moreover, the T -T dimer method has been
revealed to have high sensitivity, good reproducibility and wide
applicability.
Yoshitaka Hishikawa, Nagasaki University School of Medicine, Department
of Histology and Cell Biology, Nagasaki, 852-8523, Japan (phone +81-95849-7027; fax +81-95-849-7028); Correspondence to Masamoto Murakami,
Fujita Health University School of Medicine, Department of Pathology, Aichi 470-1192, Toyoake-cityJapan (phone +81-562-93-2440;fax +81-562-932440); Masao Kasahara, Fujita Health University School of Medicine, Department of Pathology, Aichi 470-1192, Toyoake-city, Japan (phone +81562-93-2440; fax +81-562-93-2440); Naofumi Nagasue, Shimane Medical
University, Second Department of Surgery, Izumo, 693-8501, Japan (phone
+81-853-20-2232; fax +81-853-20-2229)
In Situ Hybridization for RNA: Nonradioactive
Probe: Oligo-DNA Probe: T -T Dimer
YOSHITAKA HISHIKAWA, MASAMOTO MURAKAMI,
MASAO KASAHARA AND NAOFUMI NAGASUE
Introduction
The method using thymine-thymine (T-T) dimerized DNA as a
haptenic probe was previously reported (Koji 1988, 1989, 1990,
1996) and we have elaborated it in the following sections.
When ultraviolet rays (UV: 254nm) are exposed to the location adjacent to thymidines in the DNA strand, Pyrimidine ring
of thymidine is detached and then new skeletal structure between both thymidines is constructed. This is called T -T dimer,
which is very stable and has antigenic property. Using this method takes advantage of the fact that T -T dimer formation in DNA
is proportional to the dose ofUV irradiation (Franklin 1982) and
the frequency of occurrence of two successive thymine bases in
the DNA, and that no purification step for isolating T -T dimerized DNA is required. Moreover, the T -T dimer method has been
revealed to have high sensitivity, good reproducibility and wide
applicability.
Yoshitaka Hishikawa, Nagasaki University School of Medicine, Department
of Histology and Cell Biology, Nagasaki, 852-8523, Japan (phone +81-95849-7027; fax +81-95-849-7028); Correspondence to Masamoto Murakami,
Fujita Health University School of Medicine, Department of Pathology, Aichi 470-1192, Toyoake-cityJapan (phone +81-562-93-2440;fax +81-562-932440); Masao Kasahara, Fujita Health University School of Medicine, Department of Pathology, Aichi 470-1192, Toyoake-city, Japan (phone +81562-93-2440; fax +81-562-93-2440); Naofumi Nagasue, Shimane Medical
University, Second Department of Surgery, Izumo, 693-8501, Japan (phone
+81-853-20-2232; fax +81-853-20-2229)