Comparative analysis of different DNA extraction protocols: A fast, universal maxi-preparation of high quality plant DNA for genetic evaluation and phylogenetic studies

被引:88
作者
Csaikl, UM
Bastian, H
Brettschneider, R
Gauch, S
Meir, A
Schauerte, M
Scholz, F
Sperisen, C
Vornam, B
Ziegenhagen, B
机构
[1] Osterreich Forschungszentrum Seibersdorf GmbH, Biotechnol Unit, A-2444 Seibersdorf, Austria
[2] QIAGEN GmbH, D-40724 Hilden, Germany
[3] AMPII, Inst Gen Bot, Ctr Appl Plant Mol Biol, D-22609 Hamburg, Germany
[4] Fed Res Ctr Forestry & Forest Prod, Inst Forest Genet, D-22927 Grosshansdorf, Germany
[5] Swiss Fed Inst Forest Snow & Landscape Res, CH-8903 Birmensdorf, Switzerland
[6] Univ Gottingen, Inst Forest Genet & Plant Breeding, D-37077 Gottingen, Germany
基金
奥地利科学基金会;
关键词
cpDNA; DNA extraction; fingerprinting; forest trees; M13; method; PCR; rDNA; RFLP; rhododendron; plant;
D O I
10.1023/A:1007428009556
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Four DNA extraction protocols were compared for ability to produce DNA from the leaves or needles of several species: oak. elm, pine, fir, poplar and maize (fresh materials) and rhododendron (silica dried or frozen material). With the exception of maize and poplar, the species are known to be difficult for DNA extraction. Two protocols represented classical procedures for lysis and purification, and the other two were a combination of classical lysis followed by anion exchange chromatography. The DNA obtained fi um all procedures was quantified and rested by PCR and Southern hybridisation. Test results indicated superiority of one of the four protocols; a combination of CTAB lysis followed by anion er;change chromatography which enabled DNA extraction from all seven species, A second protocol also produced DNA from leaves or needles of all species investigated and was well suited for PCR applications but not Southern hybridisations. The remaining protocols produced DNA from some but not all species tested.
引用
收藏
页码:69 / 86
页数:18
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