Direct visualization of single copy genes on banded metaphase chromosomes by nonisotopic in situ hybridization
A rapid method is described for non isotopic in situ mapping of single copy genes directly on G-banded chromosomes by "one-step" regular light microscopy. It is based on hybridizing biotinylated probes to metaphase chromosomes. Biotin residues are detected by rabbit antibiotin antibody and...
Gespeichert in:
| Veröffentlicht in: | Nucleic acids research 1988-05, Vol.16 (9), p.3951-3961 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 3961 |
|---|---|
| container_issue | 9 |
| container_start_page | 3951 |
| container_title | Nucleic acids research |
| container_volume | 16 |
| creator | BHUPENDRA BHATT BURNS, J FLANNERY, D MCGEE, J. O'D |
| description | A rapid method is described for non isotopic in situ mapping of single copy genes directly on G-banded chromosomes by "one-step" regular light microscopy. It is based on hybridizing biotinylated probes to metaphase chromosomes. Biotin residues are detected by rabbit antibiotin antibody and anti-rabbit Ig labelled with peroxidase or colloidal gold. The peroxidase reaction product or colloidal gold signals are amplified by silver precipitation. The final product is a black silver dot at the gene locus on a purple G-banded chromosome. N-ras and alpha-1-antitrypsin genes have been mapped using plasmids with inserts of 1.5 and 1.3kb to 1p13.1 and the junction of 14q31/32 respectively. The signal to noise ratio in these experiments ranged from 32:1-46:1. This technology is at least as sensitive as radioisotopic in situ hybridization and gives results within 1 day of hybridization and has much better resolution. Additionally, genes are visualized by regular light microscopy without specialized techniques such as reflection contrast, fluorescence or phase microscopy. This methodology should facilitate more precise chromosomal gene localization. |
| doi_str_mv | 10.1093/nar/16.9.3951 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_336567</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>20207873</sourcerecordid><originalsourceid>FETCH-LOGICAL-c348t-89360ef421812828f22c334d91a107105a8a03e835c76e587a81a1901cb368df3</originalsourceid><addsrcrecordid>eNqFkU1rFTEUhoMo9ba6dClkIe7mNsnJ1yxcSNUqFNzoOmQymXsjM8mYzBSuv96UDhdddXUgz3NeTngRekPJnpIWrqPN11Tu2z20gj5DOwqSNbyV7DnaESCioYTrl-iylF-EUE4Fv0AXAEoQJXYofgrZuwXfh7LaMfyxS0gRpwGXEA-jxy7NJ3zw0Rdc3zsbe9_jyS92PtpS8TGnKZU0Vd6dcEwxlLSkOTgcYs1YVnw8dTn0W_Ir9GKwY_Gvt3mFfn75_OPma3P3_fbbzce7xgHXS6NbkMQPnFFNmWZ6YMwB8L6llhJFibDaEvAahFPSC62srqQl1HUgdT_AFfrwmDuv3eR75-OS7WjmHCabTybZYP4nMRzNId0bACmkqvvvt_2cfq--LGYKxflxtNGntRilGSjFyJMiI4woreBJkQpCtOS8is2j6HIqJfvhfDUl5qFxUxs3VJrWPDRe_bf_fvVsbxVX_m7jtjg7DtlGF8pZU5ILIQH-AsrNtdA</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>15008644</pqid></control><display><type>article</type><title>Direct visualization of single copy genes on banded metaphase chromosomes by nonisotopic in situ hybridization</title><source>MEDLINE</source><source>PubMed Central</source><source>Oxford University Press Journals Digital Archive Legacy</source><creator>BHUPENDRA BHATT ; BURNS, J ; FLANNERY, D ; MCGEE, J. O'D</creator><creatorcontrib>BHUPENDRA BHATT ; BURNS, J ; FLANNERY, D ; MCGEE, J. O'D</creatorcontrib><description>A rapid method is described for non isotopic in situ mapping of single copy genes directly on G-banded chromosomes by "one-step" regular light microscopy. It is based on hybridizing biotinylated probes to metaphase chromosomes. Biotin residues are detected by rabbit antibiotin antibody and anti-rabbit Ig labelled with peroxidase or colloidal gold. The peroxidase reaction product or colloidal gold signals are amplified by silver precipitation. The final product is a black silver dot at the gene locus on a purple G-banded chromosome. N-ras and alpha-1-antitrypsin genes have been mapped using plasmids with inserts of 1.5 and 1.3kb to 1p13.1 and the junction of 14q31/32 respectively. The signal to noise ratio in these experiments ranged from 32:1-46:1. This technology is at least as sensitive as radioisotopic in situ hybridization and gives results within 1 day of hybridization and has much better resolution. Additionally, genes are visualized by regular light microscopy without specialized techniques such as reflection contrast, fluorescence or phase microscopy. This methodology should facilitate more precise chromosomal gene localization.</description><identifier>ISSN: 0305-1048</identifier><identifier>EISSN: 1362-4962</identifier><identifier>DOI: 10.1093/nar/16.9.3951</identifier><identifier>PMID: 3375075</identifier><identifier>CODEN: NARHAD</identifier><language>eng</language><publisher>Oxford: Oxford University Press</publisher><subject>Biological and medical sciences ; Cells, Cultured ; Chromosome Banding ; Chromosomes, Human - ultrastructure ; Fundamental and applied biological sciences. Psychology ; Genes ; Genes. Genome ; Humans ; Karyotyping ; Lymphocytes - cytology ; Metaphase ; Mitosis ; Molecular and cellular biology ; Molecular genetics ; Nucleic Acid Hybridization</subject><ispartof>Nucleic acids research, 1988-05, Vol.16 (9), p.3951-3961</ispartof><rights>1988 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c348t-89360ef421812828f22c334d91a107105a8a03e835c76e587a81a1901cb368df3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC336567/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC336567/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7645563$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3375075$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>BHUPENDRA BHATT</creatorcontrib><creatorcontrib>BURNS, J</creatorcontrib><creatorcontrib>FLANNERY, D</creatorcontrib><creatorcontrib>MCGEE, J. O'D</creatorcontrib><title>Direct visualization of single copy genes on banded metaphase chromosomes by nonisotopic in situ hybridization</title><title>Nucleic acids research</title><addtitle>Nucleic Acids Res</addtitle><description>A rapid method is described for non isotopic in situ mapping of single copy genes directly on G-banded chromosomes by "one-step" regular light microscopy. It is based on hybridizing biotinylated probes to metaphase chromosomes. Biotin residues are detected by rabbit antibiotin antibody and anti-rabbit Ig labelled with peroxidase or colloidal gold. The peroxidase reaction product or colloidal gold signals are amplified by silver precipitation. The final product is a black silver dot at the gene locus on a purple G-banded chromosome. N-ras and alpha-1-antitrypsin genes have been mapped using plasmids with inserts of 1.5 and 1.3kb to 1p13.1 and the junction of 14q31/32 respectively. The signal to noise ratio in these experiments ranged from 32:1-46:1. This technology is at least as sensitive as radioisotopic in situ hybridization and gives results within 1 day of hybridization and has much better resolution. Additionally, genes are visualized by regular light microscopy without specialized techniques such as reflection contrast, fluorescence or phase microscopy. This methodology should facilitate more precise chromosomal gene localization.</description><subject>Biological and medical sciences</subject><subject>Cells, Cultured</subject><subject>Chromosome Banding</subject><subject>Chromosomes, Human - ultrastructure</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes</subject><subject>Genes. Genome</subject><subject>Humans</subject><subject>Karyotyping</subject><subject>Lymphocytes - cytology</subject><subject>Metaphase</subject><subject>Mitosis</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Nucleic Acid Hybridization</subject><issn>0305-1048</issn><issn>1362-4962</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1988</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1rFTEUhoMo9ba6dClkIe7mNsnJ1yxcSNUqFNzoOmQymXsjM8mYzBSuv96UDhdddXUgz3NeTngRekPJnpIWrqPN11Tu2z20gj5DOwqSNbyV7DnaESCioYTrl-iylF-EUE4Fv0AXAEoQJXYofgrZuwXfh7LaMfyxS0gRpwGXEA-jxy7NJ3zw0Rdc3zsbe9_jyS92PtpS8TGnKZU0Vd6dcEwxlLSkOTgcYs1YVnw8dTn0W_Ir9GKwY_Gvt3mFfn75_OPma3P3_fbbzce7xgHXS6NbkMQPnFFNmWZ6YMwB8L6llhJFibDaEvAahFPSC62srqQl1HUgdT_AFfrwmDuv3eR75-OS7WjmHCabTybZYP4nMRzNId0bACmkqvvvt_2cfq--LGYKxflxtNGntRilGSjFyJMiI4woreBJkQpCtOS8is2j6HIqJfvhfDUl5qFxUxs3VJrWPDRe_bf_fvVsbxVX_m7jtjg7DtlGF8pZU5ILIQH-AsrNtdA</recordid><startdate>19880511</startdate><enddate>19880511</enddate><creator>BHUPENDRA BHATT</creator><creator>BURNS, J</creator><creator>FLANNERY, D</creator><creator>MCGEE, J. O'D</creator><general>Oxford University Press</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19880511</creationdate><title>Direct visualization of single copy genes on banded metaphase chromosomes by nonisotopic in situ hybridization</title><author>BHUPENDRA BHATT ; BURNS, J ; FLANNERY, D ; MCGEE, J. O'D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c348t-89360ef421812828f22c334d91a107105a8a03e835c76e587a81a1901cb368df3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1988</creationdate><topic>Biological and medical sciences</topic><topic>Cells, Cultured</topic><topic>Chromosome Banding</topic><topic>Chromosomes, Human - ultrastructure</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes</topic><topic>Genes. Genome</topic><topic>Humans</topic><topic>Karyotyping</topic><topic>Lymphocytes - cytology</topic><topic>Metaphase</topic><topic>Mitosis</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Nucleic Acid Hybridization</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>BHUPENDRA BHATT</creatorcontrib><creatorcontrib>BURNS, J</creatorcontrib><creatorcontrib>FLANNERY, D</creatorcontrib><creatorcontrib>MCGEE, J. O'D</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Nucleic acids research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>BHUPENDRA BHATT</au><au>BURNS, J</au><au>FLANNERY, D</au><au>MCGEE, J. O'D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Direct visualization of single copy genes on banded metaphase chromosomes by nonisotopic in situ hybridization</atitle><jtitle>Nucleic acids research</jtitle><addtitle>Nucleic Acids Res</addtitle><date>1988-05-11</date><risdate>1988</risdate><volume>16</volume><issue>9</issue><spage>3951</spage><epage>3961</epage><pages>3951-3961</pages><issn>0305-1048</issn><eissn>1362-4962</eissn><coden>NARHAD</coden><abstract>A rapid method is described for non isotopic in situ mapping of single copy genes directly on G-banded chromosomes by "one-step" regular light microscopy. It is based on hybridizing biotinylated probes to metaphase chromosomes. Biotin residues are detected by rabbit antibiotin antibody and anti-rabbit Ig labelled with peroxidase or colloidal gold. The peroxidase reaction product or colloidal gold signals are amplified by silver precipitation. The final product is a black silver dot at the gene locus on a purple G-banded chromosome. N-ras and alpha-1-antitrypsin genes have been mapped using plasmids with inserts of 1.5 and 1.3kb to 1p13.1 and the junction of 14q31/32 respectively. The signal to noise ratio in these experiments ranged from 32:1-46:1. This technology is at least as sensitive as radioisotopic in situ hybridization and gives results within 1 day of hybridization and has much better resolution. Additionally, genes are visualized by regular light microscopy without specialized techniques such as reflection contrast, fluorescence or phase microscopy. This methodology should facilitate more precise chromosomal gene localization.</abstract><cop>Oxford</cop><pub>Oxford University Press</pub><pmid>3375075</pmid><doi>10.1093/nar/16.9.3951</doi><tpages>11</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0305-1048 |
| ispartof | Nucleic acids research, 1988-05, Vol.16 (9), p.3951-3961 |
| issn | 0305-1048 1362-4962 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_336567 |
| source | MEDLINE; PubMed Central; Oxford University Press Journals Digital Archive Legacy |
| subjects | Biological and medical sciences Cells, Cultured Chromosome Banding Chromosomes, Human - ultrastructure Fundamental and applied biological sciences. Psychology Genes Genes. Genome Humans Karyotyping Lymphocytes - cytology Metaphase Mitosis Molecular and cellular biology Molecular genetics Nucleic Acid Hybridization |
| title | Direct visualization of single copy genes on banded metaphase chromosomes by nonisotopic in situ hybridization |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-05T14%3A18%3A09IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Direct%20visualization%20of%20single%20copy%20genes%20on%20banded%20metaphase%20chromosomes%20by%20nonisotopic%20in%20situ%20hybridization&rft.jtitle=Nucleic%20acids%20research&rft.au=BHUPENDRA%20BHATT&rft.date=1988-05-11&rft.volume=16&rft.issue=9&rft.spage=3951&rft.epage=3961&rft.pages=3951-3961&rft.issn=0305-1048&rft.eissn=1362-4962&rft.coden=NARHAD&rft_id=info:doi/10.1093/nar/16.9.3951&rft_dat=%3Cproquest_pubme%3E20207873%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=15008644&rft_id=info:pmid/3375075&rfr_iscdi=true |