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Improvement of in situ PCR by optimization of PCR cycle number and proteinase k concentration: localization of x chromosome-linked phosphoglycerate kinase-1 gene in mouse reproductive organs.
http://hdl.handle.net/10069/23714
http://hdl.handle.net/10069/237140f3927ee-bc30-4a20-bcd1-c9c0b06aea10
名前 / ファイル | ライセンス | アクション |
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AHC42_15.pdf (1.2 MB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2010-08-31 | |||||
タイトル | ||||||
タイトル | Improvement of in situ PCR by optimization of PCR cycle number and proteinase k concentration: localization of x chromosome-linked phosphoglycerate kinase-1 gene in mouse reproductive organs. | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | in situ PCR | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | pgk-1 | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | ovary | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | testis | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | proteinase K | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者 |
Hishikawa, Yoshitaka
× Hishikawa, Yoshitaka× An, Shucai× Yamamoto-Fukuda, Tomomi× Shibata, Yasuaki× Koji, Takehiko |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | In situ polymerase chain reaction (in situ PCR), which can detect a few copies of genes within a cell by amplifying the target gene, was developed to better understand the biological functions of tissues. In this study, we optimized the protocol conditions for the detection of X chromosome-linked phosphoglycerate kinase-1 (pgk-1) gene in paraffin-embedded sections of mouse reproductive organs. The effects of various concentrations of proteinase K (PK) and PCR cycle numbers were examined. To label the amplified DNA, we used digoxigenin-dUTP (Dig), Cy-3-dUTP (Cy-3), or FluorX-dCTP (FluorX). The optimal concentration of PK was 50 microg/ml for the ovary and 10 microg/ml for the testis. Ten PCR cycles were optimal for Dig and 25 cycles were optimal for FluorX and Cy-3 in the ovary and testis. The signal-to-noise ratio of FluorX and Cy-3 for ovarian tissue was better than that of Dig. Using the above conditions, we detected 1-4 and 1-2 spots of pgk-1 in the nuclei of granulosa and germ cells, respectively. Our results indicate that in situ PCR is useful for detecting a specific gene in paraffin-embedded sections under optimized conditions of both PCR cycle number and PK concentration. | |||||
書誌情報 |
Acta histochemica et cytochemica 巻 42, 号 2, p. 15-21, 発行日 2009-04-28 |
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出版者 | ||||||
出版者 | The Japan Society of Histochemistry and Cytochemistry | |||||
出版者別言語 | ||||||
日本組織細胞化学会 | ||||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 00445991 | |||||
EISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 13475800 | |||||
書誌レコードID | ||||||
収録物識別子タイプ | NCID | |||||
収録物識別子 | AA00508022 | |||||
PubMed番号 | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | PMID | |||||
関連識別子 | 19492023 | |||||
DOI | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | DOI | |||||
関連識別子 | 10.1267/ahc.09011 | |||||
権利 | ||||||
権利情報 | Copyright ©2009 By the Japan Society of Histochemistry and Cytochemistry | |||||
著者版フラグ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||
引用 | ||||||
内容記述タイプ | Other | |||||
内容記述 | Acta Histochemica et Cytochemica, 42(2), pp.15-21; 2009 |