Subscribe Now Subscribe Today
Science Alert
 
FOLLOW US:     Facebook     Twitter
Blue
   
Curve Top
International Journal of Botany
  Year: 2010 | Volume: 6 | Issue: 3 | Page No.: 293-298
DOI: 10.3923/ijb.2010.293.298
 
Facebook Twitter Digg Reddit Linkedin StumbleUpon E-mail
Standardization of DNA Extraction and Optimization of RAPD-PCR Conditions in Garcinia indica
Abhijit Sahasrabudhe and Manjushree Deodhar

Abstract:
Genetic analysis of plant relies on high yields of pure DNA samples. DNA isolation is difficult in woody plants because of presence of polysaccharides, tannins, alkaloids, polyphenols and other secondary metabolites that interfere during isolation. Here we report for the first time a fast, reliable and less expensive method of genomic DNA isolation from leaves of Garcinia indica. The modified CTAB protocol includes addition of Polyvinylpyrrolidone (PVP) separately in each tube and precipitation with 5M NaCl along with chilled ethanol, which increased the solubility of polysaccharides. Without use of RNase or Proteinase K, were able to isolate pure and sufficient amount of DNA, which proved to be amenable to RAPD analysis. We also optimized RAPD-PCR conditions such as annealing temp, amount of DNA and Taq polymerase etc. A preliminary study of variation within G. indica species was carried out with nine plants with twenty decamers. Out of which six primers showed polymorphism while three had monomorphic banding pattern. In continuation with these promising results, efforts are underway to screen more plants with different geographical locations and with more number of primers to study genetic diversity.
PDF Fulltext XML References Citation Report Citation
 RELATED ARTICLES:
  •    Optimization of DNA Extraction Methods from Garcinia species for ISSR-PCR, RAPD-PCR and DNA Barcoding
  •    Determination of Quantity and Purity of Some Geophytes DNA Collected from the Flora of Turkey
  •    Development of Multiplex PCR (Polymerase Chain Reaction) Method for Detection of Salmonella spp. and Vibrio parahaemolyticus from Shrimp Samples of Bangladesh
  •    Optimization of DNA Isolation and PCR Protocol for ISSR Analysis of Nothapodytes nimmoniana: A Threatened Anti-cancerous Medicinal Plant
How to cite this article:

Abhijit Sahasrabudhe and Manjushree Deodhar, 2010. Standardization of DNA Extraction and Optimization of RAPD-PCR Conditions in Garcinia indica. International Journal of Botany, 6: 293-298.

DOI: 10.3923/ijb.2010.293.298

URL: https://scialert.net/abstract/?doi=ijb.2010.293.298

 
COMMENT ON THIS PAPER
 
 
 

 

 
 
 
 
 
 
 
 
 

 
 
 
 
 
 
 
 
 
 

       

       

Curve Bottom