Rapid, Effective DNA Isolation from Osmanthus via Modified Alkaline Lysis

Area of Science:

• Plant molecular biology
• Genetics
• Biotechnology

Background:

• Osmanthus species present challenges for DNA extraction due to leaf structure and secondary metabolites.
• Reliable DNA isolation is essential for genetic studies and cultivar identification.

Purpose of the Study:

• To develop a universal, rapid, effective, and cost-efficient DNA isolation method for mature Osmanthus leaf tissue.
• To compare the performance of four different DNA extraction techniques.

Main Methods:

• Four DNA isolation methods were tested on 8 Osmanthus cultivars: solid-phase extraction (SPE), modified alkaline lysis (SDS), CTAB with chloroform (CHL), and CTAB with phenol/chloroform (PHE).
• DNA quality, quantity, and integrity were assessed using absorbance spectra, agarose gel electrophoresis, and PCR amplification.

Main Results:

• The modified alkaline lysis (SDS) method yielded the highest quantity of DNA (117 ± 54.1 μg) and was cost-efficient (approx. $0.74 USD/sample) and rapid (∼1.5 h).
• SPE resulted in the least contamination but lower DNA yield and higher cost.
• SDS and SPE showed better DNA integrity on agarose gels compared to CTAB methods; all methods except PHE performed well in PCR.

Conclusions:

• Improved alkaline lysis (SDS) is recommended as a rapid, effective, and cost-efficient method for isolating DNA from Osmanthus species.
• This method facilitates high-throughput DNA extraction for genetic research and breeding programs.