Separation of DNA for molecular markers analysis from leaves of the Vitis vinifera.
In the present study, three DNA extraction procedures were examined to determine which might yield DNA from Grape leaves suitable for molecular analysis for RAPD, SSR. AFLP and etc analysis. The three methods examined were: the miniprep procedure and the modified CTAB for difficult species and protocol CTAB. Only the modified CTAB method consistently yielded DNA suitable for Polymerase Chain Reaction (PCR) amplification, regardless of plant growing conditions or leaf age. The quality and quantity of extracted genomic DNA gained from these methods are deliberated by means UV biophotometer, electrophoresis in 1.2% agarose gel and PCR. In this regard, application chosen for young and mature leaves, the most value of qualified DNA, is extracted from fully expanded leave when PVP was added to the extraction buffer. This same procedure also yielded PCR-amplifiable DNA from various other perennial, woody species and from other fruit species such as apple (Malus domestica), cherry (Prunus avium), peach (Prunuspersica), plum (Prunus domestica). DNA yield from this procedure is high (up to 1 mg g(-1) of leaf tissue). DNA is completely digestible with restriction endonucleases and amplifiable in the Polymerase Chain Reaction (PCR).
Nazhad
Nafiseh Rigi
NR
Department of Plant Breeding-Biocentre, Faculty of Agriculture, Zabol University, Zabol, Iran.
Solouki
Mahmood
M
D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification*; D017931:DNA Primers; D017343:Genes, Plant; D005819:Genetic Markers; D018515:Plant Leaves / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique; D027843:Vitis / Q000737:chemistry / Q000235:genetics*
D017931:DNA Primers; D005819:Genetic Markers; D004247:DNA