Selection of chloroplasts by laser microbeam microdissection for single-chloroplast PCR

Harald Meimberg, S. Thalhammer, A. Brachmann, B. Müller, L. A. Eichacker, W. M. Heckl, G. Heubl

Publikation: Beitrag in FachzeitschriftArtikelBegutachtung

27 Zitate (Scopus)

Abstract

Laser microbeam microdissection and laser pressure catapulting offer the possibility of separating cell compartments, thus allowing for contamination-free analysis. Using these methods, we were able to select single chloroplasts of Nicotiana tabacum. Starting from homogenized leaf material, chloroplasts were purified by differential centrifugation and applied directly onto a poly-ethylene-naphthalate membrane that was mounted on a microscope slide. Single chloroplasts were dissected under microscopic control and catapulted into a PCR tube. Subsequent PCR of a spacer region between the trnT and trnF genes verified the successful amplification of DNA from a single chloroplast. The advantage of this method compared to the use of capillaries or optical tweezers is that one is able to prepare high numbers of samples in a short time.

OriginalspracheEnglisch
Seiten (von - bis)1238-1243
Seitenumfang6
FachzeitschriftBioTechniques
Jahrgang34
Ausgabenummer6
DOIs
PublikationsstatusVeröffentlicht - 1 Juni 2003
Extern publiziertJa

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