Non-cloning Synthesis of cRNA Probe and Its Application to In Situ Hybridization Analysis of Formalin-fixed Paraffin Embedded Tissue

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概要

• 論文の詳細を見る

• Intestinal type alkaline phosphatase (IALP) mRNA-specific cRNA probe was synthesized by non-cloning method for in situ hybridization (ISH) analysis. Total RNA was extracted from small intestine of male F344 rat. Reverse transcription-polymerase chain reaction (RT-PCR) was performed for IALP gene. T7 promoter adapter was directly added to IALP cDNA-specific PCR product by ligation reaction. IALP cDNA accompanied by T7 promoter adapter was then amplified by PCR using IALP gene-specific primer in combination with T7 promoter adapter primer sequence and used as a template of cRNA probe. Digoxigenin-labeled cRNA was synthesized by in vitro transcription method using RNA polimerase. ISH was performed using formalin-fixed, paraffin embedded small intestine tissue. As a result, the cytoplasm of mucosal epithelial cells was successfully stained with this cRNA probe. The present procedure is quite simple and a rapid way to prepare the cRNA probe and would be very useful for ISH analysis of formalin-fixed paraffin embedded tissue sample.

• 日本毒性病理学会の論文
• 2003-12-30

著者

• Goda Maki

  Discovery Technology Laboratory Mitsubishi Pharma Co.

• Fukunari Atsushi

  Discovery Technology Laboratory Mitsubishi Pharma Co.

• KOBAYASHI Kiyoshi

  Discovery Technology Laboratory, Mitsubishi Pharma Co.

• TANAKA Hideki

  Discovery Technology Laboratory, Mitsubishi Pharma Co.

• Kobayashi Kiyoshi

  Discovery Technology Laboratory Mitsubishi Pharma Co.

• Tanaka Hideki

  Discovery Technology Laboratory Mitsubishi Pharma Co.

関連論文

• Non-cloning Synthesis of cRNA Probe and Its Application to In Situ Hybridization Analysis of Formalin-fixed Paraffin Embedded Tissue

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