Optimizing DNA extraction procedure in case of Amygdalus spp. | ||
| تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران | ||
| Article 2, Volume 10, Issue 1 - Serial Number 10, January 2003, Pages 15-27 PDF (500.34 K) | ||
| DOI: 10.22092/ijrfpbgr.2002.115682 | ||
| Authors | ||
| s. Kadkhodaie* 1; S.R. Tabaie-Aghduie22 | ||
| 11 - Postgraduate student, Tabriz University 2 - Research Institute of Fortests and Rangelands, P.O.Box 13185-l16, Tehran | ||
| 2Research 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 | ||
| DNA extraction; Amygdalus spp; Polysaccharides; Polyphenoles | ||
| References | ||
|
clarck M. s., lggT. Plant molecular biology-A laboratory manual. | ||
|
Statistics Article View: 567 PDF Download: 447 |
||