A simple method for obtaining cell-specific cDNA from small numbers of growing root-hair cells in Arabidopsis thaliana
A simple and rapid method for cloning specific cDNAs from mRNA populations derived solely from small numbers of root-hair cells is described here. To identify genes expressed during the earliest visible stage of root-hair cell development, cell contents were aspirated from small numbers of Arabidops...
| Veröffentlicht in: | Journal of experimental botany. - 1985. - 54(2003), 386 vom: 04. Mai, Seite 1373-8 |
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| Format: | Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2003
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| Zugriff auf das übergeordnete Werk: | Journal of experimental botany |
| Schlagworte: | Journal Article Research Support, Non-U.S. Gov't Arabidopsis Proteins DNA, Complementary RNA, Messenger GTP-Binding Proteins EC 3.6.1.- RHD3 protein, Arabidopsis rho GTP-Binding Proteins EC 3.6.5.2 |
| Zusammenfassung: | A simple and rapid method for cloning specific cDNAs from mRNA populations derived solely from small numbers of root-hair cells is described here. To identify genes expressed during the earliest visible stage of root-hair cell development, cell contents were aspirated from small numbers of Arabidopsis root-hair cells at or just before this stage. This material was used to make reusable solid-phase oligo-dT-primed cDNA libraries. To demonstrate that the libraries contained high quality longer cDNAs, a fragment located 2.7 kb from the 3' end of the cDNA of the single copy root-hair expressed gene RHD3 was cloned using a nested PCR strategy. This technique was also used to obtain novel gene expression information by cloning the full-length 0.85 kb cDNA of the Rop2 GTPase from this library. This approach offers a means of cloning larger cDNAs directly from small numbers of growing root-hair cells and, potentially, other epidermal cell types |
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| Beschreibung: | Date Completed 25.07.2003 Date Revised 13.05.2019 published: Print Citation Status MEDLINE |
| ISSN: | 1460-2431 |