Improved protocol for isolation of genomic DNA from leaf tissues of Phyllanthus emblica Gaertn

Document Type: Brief Report

Authors

1 Department of Biotechnology, Plant Genetic Improvement Laboratory, Sri Paramakalyani Centre for Environmental Sciences, Manonmaniam Sundaranar University, Alwarkurichi 627 412, Tirunelveli District, Tamilnadu, India.

2 1Department of Biotechnology, Plant Genetic Improvement Laboratory, Sri Paramakalyani Centre for Environmental Sciences, Manonmaniam Sundaranar University, Alwarkurichi 627 412, Tirunelveli District, Tamilnadu, India.

3 Department of Plant Biotechnology, School of Biotechnology, Madurai Kamarai University, Palkalainagar, Madurai 625 021, India.

Abstract

Modified Cetyltrimethylammonium bromide (CTAB) protocol for DNA isolation was developed from leaf tissues
of Phyllanthus emblica for obtaining high quality genomic DNA. Fresh leaves of three different maturity
were analyzed for yield and quality of DNA. Acidity was determined in three different maturity of leaves viz.
tender, intermediate and mature and their influence on DNA quality was determined. Drastic reduction of pH
was the primary cause for poor quality of DNA. However, high quality DNA isolation was achieved by
stabilizing the pH by addition of NaOH during different stages of DNA isolation process. The present protocol
yielding high quality intact DNA for genetic fingerprinting as well as for amplification of chloroplast genes for
molecular analysis.

Keywords