Does extraction of DNA and RNA by magnetic fishing work for diverse plant species?
An automated nucleic acid extraction procedure with magnetic particles originally designed for isolation of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from animal tissues was tested for plant material. We isolated genomic DNA and total RNA from taxonomically diverse plant species representing conifers (Scots pine), broad-leaved trees (silver birch and hybrid aspen), dwarf shrubs (bilberry), and both monocotyledonous (regal lily) and dicotyledonous (Saint John's wort, round-leaved sundew, and tobacco) herbaceous plants. Buffers developed for DNA extraction were successfully used in addition to manufacturer's extraction kits. The quality of RNA was appropriate for many applications, but the quality of DNA was not always sufficient for polymerase chain reaction (PCR) amplification. However, we could strikingly improve the quality by eliminating the adherent compounds during the extraction or later in the PCR phase. Our results show that the use of the procedure could be extended to diverse plant species. This procedure is especially suitable for small sample sizes and for simultaneous processing of many samples enabling large-scale plant applications in population genetics, or in the screening of putative transgenic plants.
Vuosku
Jaana
J
Department of Biology, University of Oulu, PO Box 3000, FIN-90014 Oulu, Finland. jaana.vuosku@oulu.fi
Jaakola
Laura
L
Jokipii
Soile
S
Karppinen
Katja
K
Kämäräinen
Terttu
T
Pelkonen
Veli-Pekka
VP
Jokela
Anne
A
Sarjala
Tytti
T
Hohtola
Anja
A
Häggman
Hely
H
D005591:Chemical Fractionation / Q000379:methods*; D018744:DNA, Plant / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D008280:Magnetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D018749:RNA, Plant / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic; D013045:Species Specificity
D018744:DNA, Plant; D018749:RNA, Plant; D011933:Reagent Kits, Diagnostic