Determination of gene copy number and genotype of transgenic Arabidopsis thaliana by competitive PCR
A simple and rapid method is described for determining the integrated T-DNA copy number and the genotype in transgenic Arabidopsis thaliana by two-step competitive PCR. First, the amount of genomic DNA in the extracts, obtained from an individual A. thaliana transformant, was accurately determined b...
| Publié dans: | Journal of experimental botany. - 1985. - 53(2002), 373 vom: 02. Juni, Seite 1515-20 |
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| Auteur principal: | |
| Autres auteurs: | , , , , |
| Format: | Article |
| Langue: | English |
| Publié: |
2002
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| Accès à la collection: | Journal of experimental botany |
| Sujets: | Comparative Study Journal Article DNA, Bacterial DNA, Plant T-DNA 4-Hydroxyphenylpyruvate Dioxygenase EC 1.13.11.27 Citrate (si)-Synthase EC 2.3.3.1 |
| Résumé: | A simple and rapid method is described for determining the integrated T-DNA copy number and the genotype in transgenic Arabidopsis thaliana by two-step competitive PCR. First, the amount of genomic DNA in the extracts, obtained from an individual A. thaliana transformant, was accurately determined by the 1st competitive PCR using a known single copy gene, 4HPPD (4-hydroxyphenylpyruvate dioxygenase), as a target. Second, the number of T-DNA copies per genome was estimated by quantifying the NPTII gene, which was involved in the T-DNA, by the 2nd competitive PCR using exactly the same amount of genomic DNA for each sample. The estimated copy number and genotype obtained by this procedure were identical to those determined by Southern blot analysis and segregation analysis |
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| Description: | Date Completed 06.09.2002 Date Revised 15.11.2006 published: Print Citation Status MEDLINE |
| ISSN: | 1460-2431 |