Chapter 8
In Situ Hybridization for RNA: Radioactive RNA
Probe - A Protocol for Practice With a Kit
HIDEFUMI YOSHIOKA AND SUMIHARE NOlI
Introduction
Recently, nonradioactive probe has often been used to detect
RNAs in sectioned tissues by in situ hybridization (ISH) method.
Although color-based detection systems with nonradioactive
probe is becoming popular in ISH methods, the method with
radioactive (RI) probes is still the most sensitive way to detect
RNAs in the tissues. Thus, if a given mRNA cannot be detected in
tissues with color-based detection methods, although it is confirmed to be present by PCR, the final way to detect the mRNA is
to use the ISH method with an RI probe. Processes of specimen
preparation, pre-treatments, and hybridization are similar in
either radioactive or nonradioactive methods. A major difference between them is the detection method, i.e., autoradiography is used for detection of an RI probe.
An efficient and easy way to learn the methods is to use a kit
including tissue sections on glass slides for a positive control,
solutions necessary for ISH, probe preparation, and detection.
We developed an ISH kit for practice, which is now commercially
available from Nippon Gene Co .. The kit has been used successfully for various tissues at least from human, mouse, rat, and
chick. In this section, the ISH kit and how to use it is introduced
only in the case of using radioactive probes and paraffin sections.
Additional information for using nonradioactive probes is also
available in the kit.
Hidefumi Yoshioka, Correspondence to Sumihare Noji, The University of
Tokushima, Department of Biological Science and Technology, Faculty of
Engineering, 2-1 Minami-Jyosanjima-cho, Tokushima City, 770-8506, Japan (phone +81-88-656-7528; fax +81-88-656-9074; e-mail noji@bio.tokushima-u.ac.jp)
In Situ Hybridization for RNA: Radioactive RNA
Probe - A Protocol for Practice With a Kit
HIDEFUMI YOSHIOKA AND SUMIHARE NOlI
Introduction
Recently, nonradioactive probe has often been used to detect
RNAs in sectioned tissues by in situ hybridization (ISH) method.
Although color-based detection systems with nonradioactive
probe is becoming popular in ISH methods, the method with
radioactive (RI) probes is still the most sensitive way to detect
RNAs in the tissues. Thus, if a given mRNA cannot be detected in
tissues with color-based detection methods, although it is confirmed to be present by PCR, the final way to detect the mRNA is
to use the ISH method with an RI probe. Processes of specimen
preparation, pre-treatments, and hybridization are similar in
either radioactive or nonradioactive methods. A major difference between them is the detection method, i.e., autoradiography is used for detection of an RI probe.
An efficient and easy way to learn the methods is to use a kit
including tissue sections on glass slides for a positive control,
solutions necessary for ISH, probe preparation, and detection.
We developed an ISH kit for practice, which is now commercially
available from Nippon Gene Co .. The kit has been used successfully for various tissues at least from human, mouse, rat, and
chick. In this section, the ISH kit and how to use it is introduced
only in the case of using radioactive probes and paraffin sections.
Additional information for using nonradioactive probes is also
available in the kit.
Hidefumi Yoshioka, Correspondence to Sumihare Noji, The University of
Tokushima, Department of Biological Science and Technology, Faculty of
Engineering, 2-1 Minami-Jyosanjima-cho, Tokushima City, 770-8506, Japan (phone +81-88-656-7528; fax +81-88-656-9074; e-mail noji@bio.tokushima-u.ac.jp)