Journal
ARCHIVES OF BIOLOGICAL SCIENCES
Volume 64, Issue 1, Pages 191-199Publisher
INST BIOLOSKA ISTRAZIVANJA SINISA STANKOVIC
DOI: 10.2298/ABS1201191S
Keywords
Centaurium; RAPD; optimization; DNA; reproducibility
Categories
Funding
- Ministry of Education and Science of the Republic of Serbia [173024]
Ask authors/readers for more resources
Random amplified polymorphic DNA (RAPD) analysis is a simple and reliable method used to detect DNA polymorphism. Several factors can affect the amplification profiles, thereby causing false bands and non-reproducibility of the assay. In this study, we analyzed the effects of different concentrations of primer, magnesium chloride, template DNA and Taq DNA polymerase to develop and standardize a RAPD protocol for Centaurium species. The optimized PCR reaction mixture included: 50 ng of DNA extracted using a CTAB based protocol, 2.5 mM MgCl2, 7.5 pmol primer and 2 U of Taq polymerase in a final volume of 25 mu l. Each of the five primers used in experiments (OPB11, OPB15, OPB18, OPF05 and OPH02) generated reproducible and distinguishable fingerprinting patterns of four Centaurium species. The obtained optimized RAPD protocol and the selected primers are useful for our further work in the genetic diversity studies of Centaurium species.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available