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JXB Advance Access originally published online on March 31, 2003
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Journal of Experimental Botany, Vol. 54, No. 386, pp. 1373-1378, May 1, 2003
© 2003 Oxford University Press

A simple method for obtaining cell-specific cDNA from small numbers of growing root-hair cells in Arabidopsis thaliana

Received 30 October 2002; Accepted 25 February 2003

Mark A. Jones1 and Claire S. Grierson

School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, UK

1 Present address and to whom correspondence should be sent: School of Biological Sciences, University of Exeter, Washington Singer Laboratories, Perry Road, Exeter, EX4 4QG, UK. Fax: +44 (0)1392 264668. E-mail: m.a.jones{at}ex.ac.uk

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.

Key words: Cell aspiration, complementary DNA, messenger RNA, nested PCR, root hairs.


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