{"created":"2023-05-15T16:41:12.932201+00:00","id":15776,"links":{},"metadata":{"_buckets":{"deposit":"e5bd730d-d9bb-4ba9-abf5-f3b5cc07b400"},"_deposit":{"created_by":2,"id":"15776","owners":[2],"pid":{"revision_id":0,"type":"depid","value":"15776"},"status":"published"},"_oai":{"id":"oai:nagasaki-u.repo.nii.ac.jp:00015776","sets":["29:30"]},"author_link":["58843","58844","58846","58845","58847"],"item_2_biblio_info_6":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2009-04-28","bibliographicIssueDateType":"Issued"},"bibliographicIssueNumber":"2","bibliographicPageEnd":"21","bibliographicPageStart":"15","bibliographicVolumeNumber":"42","bibliographic_titles":[{"bibliographic_title":"Acta histochemica et cytochemica"}]}]},"item_2_description_4":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"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.","subitem_description_type":"Abstract"}]},"item_2_description_63":{"attribute_name":"引用","attribute_value_mlt":[{"subitem_description":"Acta Histochemica et Cytochemica, 42(2), pp.15-21; 2009","subitem_description_type":"Other"}]},"item_2_publisher_33":{"attribute_name":"出版者","attribute_value_mlt":[{"subitem_publisher":"The Japan Society of Histochemistry and Cytochemistry"}]},"item_2_relation_11":{"attribute_name":"PubMed番号","attribute_value_mlt":[{"subitem_relation_type":"isIdenticalTo","subitem_relation_type_id":{"subitem_relation_type_id_text":"19492023","subitem_relation_type_select":"PMID"}}]},"item_2_relation_12":{"attribute_name":"DOI","attribute_value_mlt":[{"subitem_relation_type":"isIdenticalTo","subitem_relation_type_id":{"subitem_relation_type_id_text":"10.1267/ahc.09011","subitem_relation_type_select":"DOI"}}]},"item_2_rights_13":{"attribute_name":"権利","attribute_value_mlt":[{"subitem_rights":"Copyright ©2009 By the Japan Society of Histochemistry and Cytochemistry"}]},"item_2_source_id_10":{"attribute_name":"書誌レコードID","attribute_value_mlt":[{"subitem_source_identifier":"AA00508022","subitem_source_identifier_type":"NCID"}]},"item_2_source_id_7":{"attribute_name":"ISSN","attribute_value_mlt":[{"subitem_source_identifier":"00445991","subitem_source_identifier_type":"ISSN"}]},"item_2_source_id_8":{"attribute_name":"EISSN","attribute_value_mlt":[{"subitem_source_identifier":"13475800","subitem_source_identifier_type":"ISSN"}]},"item_2_text_62":{"attribute_name":"出版者別言語","attribute_value_mlt":[{"subitem_text_value":"日本組織細胞化学会"}]},"item_2_version_type_16":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"Hishikawa, Yoshitaka"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"An, Shucai"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Yamamoto-Fukuda, Tomomi"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Shibata, Yasuaki"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Koji, Takehiko"}],"nameIdentifiers":[{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2020-12-23"}],"displaytype":"detail","filename":"AHC42_15.pdf","filesize":[{"value":"1.2 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"AHC42_15.pdf","url":"https://nagasaki-u.repo.nii.ac.jp/record/15776/files/AHC42_15.pdf"},"version_id":"603b2561-75c4-41b5-8c5b-e37f8afcf8c8"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"in situ PCR","subitem_subject_scheme":"Other"},{"subitem_subject":"pgk-1","subitem_subject_scheme":"Other"},{"subitem_subject":"ovary","subitem_subject_scheme":"Other"},{"subitem_subject":"testis","subitem_subject_scheme":"Other"},{"subitem_subject":"proteinase K","subitem_subject_scheme":"Other"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"journal article","resourceuri":"http://purl.org/coar/resource_type/c_6501"}]},"item_title":"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.","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"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."}]},"item_type_id":"2","owner":"2","path":["30"],"pubdate":{"attribute_name":"公開日","attribute_value":"2010-08-31"},"publish_date":"2010-08-31","publish_status":"0","recid":"15776","relation_version_is_last":true,"title":["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."],"weko_creator_id":"2","weko_shared_id":2},"updated":"2023-05-15T19:08:58.453651+00:00"}