Mass cloning of differential and nondifferential transcript-derived fragments from cDNA-AFLP experiments in sunflower
We have developed a 1-step procedure for cloning multiple bands obtained through cDNA-amplified fragment length polymorphism (AFLP) experiments. This protocol enables us to clone differential and nondifferential transcript-derived fragments (TDFs) in the same experiment. It consists of a mass clonin...
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| Veröffentlicht in: | Plant molecular biology reporter 2004-06, Vol.22 (2), p.165-171 |
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| creator | Tamborindeguy,C Ben, C Jardinaud, F Gentzbittel, L Liboz, T |
| description | We have developed a 1-step procedure for cloning multiple bands obtained through cDNA-amplified fragment length polymorphism (AFLP) experiments. This protocol enables us to clone differential and nondifferential transcript-derived fragments (TDFs) in the same experiment. It consists of a mass cloning of cDNA-AFLP amplification products, identification of clones of interest with electrophoresis, and characterization through sequencing. The entire experiment can be performed in 1 wk because of the reduced time and effort needed for cloning, and it can be applied to any PCR method that produces complex band patterns. This method, tested on a comparison of gene expression between globular and heart embryo stages in sunflower embryogenesis, enabled us to clone 10 differential TDFs in the same experiment. |
| doi_str_mv | 10.1007/BF02772723 |
| format | Article |
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This protocol enables us to clone differential and nondifferential transcript-derived fragments (TDFs) in the same experiment. It consists of a mass cloning of cDNA-AFLP amplification products, identification of clones of interest with electrophoresis, and characterization through sequencing. The entire experiment can be performed in 1 wk because of the reduced time and effort needed for cloning, and it can be applied to any PCR method that produces complex band patterns. 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This method, tested on a comparison of gene expression between globular and heart embryo stages in sunflower embryogenesis, enabled us to clone 10 differential TDFs in the same experiment.</description><subject>Amplified fragment length polymorphism</subject><subject>Cloning</subject><subject>cloning (plants)</subject><subject>complementary DNA</subject><subject>Embryogenesis</subject><subject>Embryonic growth stage</subject><subject>Experiments</subject><subject>Fragmentation</subject><subject>Fragments</subject><subject>Gene expression</subject><subject>Helianthus annuus</subject><subject>oilseed crops</subject><subject>Polymorphism</subject><subject>Transcription</subject><subject>transcription (genetics)</subject><subject>transgenic plants</subject><issn>0735-9640</issn><issn>1572-9818</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNpVkFtLAzEQhYMoWKsv_gEDvgmrSTa7SR7rpSrUC2ifl2l2UrZsk5psvfx7V1YQn2aY83HOcAg55uycM6YuLqdMKCWUyHfIiBdKZEZzvUtGTOVFZkrJ9slBSivWw0zrEdk-QErUtsE3fkmDo3XjHEb0XQMtBV9TH_y_WxfBJxubTZfVGJt3rKmLsFz3cuq3sKb2-nGSTaazZ4qfmx4ZpMbTtPWuDR8YD8megzbh0e8ck_n05vXqLps93d5fTWaZFTrvMrWQCg04LFUBRqKueS1q64A7BFBFyWrkqBGkNTlqCVJwo1E4i7aEhcnH5HTw3cTwtsXUVauwjb6PrLgpZCllzsqeOhsoG0NKEV216Z-G-FVxVv3UWv3V2sMnA-wgVLCMTarmL4LxnDFTmLIo82_V_XX3</recordid><startdate>20040601</startdate><enddate>20040601</enddate><creator>Tamborindeguy,C</creator><creator>Ben, C</creator><creator>Jardinaud, F</creator><creator>Gentzbittel, L</creator><creator>Liboz, T</creator><general>Springer Nature B.V</general><scope>FBQ</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7QR</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FK</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>LK8</scope><scope>M0K</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope></search><sort><creationdate>20040601</creationdate><title>Mass cloning of differential and nondifferential transcript-derived fragments from cDNA-AFLP experiments in sunflower</title><author>Tamborindeguy,C ; 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| subjects | Amplified fragment length polymorphism Cloning cloning (plants) complementary DNA Embryogenesis Embryonic growth stage Experiments Fragmentation Fragments Gene expression Helianthus annuus oilseed crops Polymorphism Transcription transcription (genetics) transgenic plants |
| title | Mass cloning of differential and nondifferential transcript-derived fragments from cDNA-AFLP experiments in sunflower |
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