Optimizing DNA extraction procedure in case of Amygdalus spp.

Authors

1 1 - Postgraduate student, Tabriz University 2 - Research Institute of Fortests and Rangelands, P.O.Box 13185-l16, Tehran

2 Research Institute of Fortests and Rangelands, P.O.Box 13185-l16, Tehran

Abstract

A simple method was developed to extract sufficient amount of genomic
DNA, from plant species containing high amount of secondary metabolites.
Leaves of different Amygdalus species including A. scoparia, A.
haussknechtii, A. lyciodes, A. elaegnifolia and A. commonis were used for
DNA extraction. SDS (instead of CTAB or Sarkosile), NaCl (to remove
polysaccharides), PVP (to remove polyphenolic compounds) were used in
extracting buffer. Sodium chloride was used again for more polysaccharide
removal and RNase to remove RNA. Also, extraction was repeated several
times, using chloform: isoamyl-alchohol to abtain more purified DNA.
Average yield of DNA, extracted with this procedure was 50 pg per I g
leaf tissue (70pglg and 30pg/g in young and old leaf tissues, respectively).
Extracted DNAs showed successful reproducibility through PCR
amplification. This method which is relativerly inexpensive, could be
efficiently applied for other species of rosaceae or different plant families
from which DNA extraction is cumbersome.

Keywords

References

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