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Titolo:
A simple and reliable method for SSU rRNA gene DNA extraction, amplification, and cloning from single AM fungal spores
Autore:
Schwarzott, D; Schussler, A;
Indirizzi:
Tech Univ Darmstadt, Inst Bot, D-64287 Darmstadt, Germany Tech Univ Darmstadt Darmstadt Germany D-64287 D-64287 Darmstadt, Germany
Titolo Testata:
MYCORRHIZA
fascicolo: 4, volume: 10, anno: 2001,
pagine: 203 - 207
SICI:
0940-6360(200101)10:4<203:ASARMF>2.0.ZU;2-J
Fonte:
ISI
Lingua:
ENG
Soggetto:
ARBUSCULAR MYCORRHIZAL FUNGI; GLOMUS-MOSSEAE; GIGASPORA-MARGARITA; GLOMALES; PCR; RDNA; IDENTIFICATION; ROOTS; DIVERSITY; SOIL;
Keywords:
DNA extraction; glomales; single spores; SSU rRNA; TOPO cloning;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Citazioni:
36
Recensione:
Indirizzi per estratti:
Indirizzo: Schussler, A Tech Univ Darmstadt, Inst Bot, Schnittspahnstr 10, D-64287 Darmstadt, Germany Tech Univ Darmstadt Schnittspahnstr 10 Darmstadt Germany D-64287
Citazione:
D. Schwarzott e A. Schussler, "A simple and reliable method for SSU rRNA gene DNA extraction, amplification, and cloning from single AM fungal spores", MYCORRHIZA, 10(4), 2001, pp. 203-207

Abstract

We describe a method that allows quick and easy PCR amplification and cloning of nearly complete SSU rRNA genes from arbuscular mycorrhizal fungi. The procedure tested on spores from 37 different glomalean isolates was basedon magnetic separation with Dynabeads, followed by nested PCR with two primer pairs. All trials led to visible amplification products of the expectedsize. Thereafter, the PCR fragments could be quickly and efficiently cloned by means of a topoi somerase-activated vector (pCR2.1-TOPO). The technique is rapid, uncomplicated and comparatively inexpensive. The use of single spores for DNA extraction has some advantages over multispore-preparations,e.g. it is less susceptible to contamination with other organisms present in the cultures. The method can be used for the quick and reliable preparation of a large number of samples and is highly reproducible. It could also be used for genes other than the SSU rRNA gene.

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Documento generato il 18/09/20 alle ore 16:49:41