Related Experiment Video
Updated: Sep 8, 2025
11:30
Automating Citrus Budwood Processing for Downstream Pathogen Detection Through Instrument Engineering
Published on: April 21, 2023
856
Improved genomic DNA extraction from citrus species using a modified CTAB method
Komaljeet Gill1, Shivanti Negi2, Pankaj Kumar3
1Department of Biotechnology, Dr. Yashwant Singh Parmar University of Horticulture and Forestry, Solan, Himachal Pradesh, India. singh.komal2111@gmail.com.
Molecular Biology Reports
|June 26, 2025
Summary
This study optimized a CTAB DNA extraction protocol for citrus, yielding high-purity genomic DNA. The improved method significantly increases DNA yield and quality for downstream applications like PCR and sequencing.
Area of Science:
- Plant Molecular Biology
- Biotechnology
- Genomics
Background:
- Citrus DNA extraction is challenging due to polysaccharides and polyphenols.
- These compounds inhibit DNA yield and purity.
- A robust protocol is needed for citrus genetic studies.
Purpose of the Study:
- To optimize a CTAB-based protocol for efficient genomic DNA isolation from citrus leaves.
- To improve DNA yield and purity compared to conventional methods.
- To provide a reliable method for citrus genetic research.
Main Methods:
- Optimized CTAB buffer with β-mercaptoethanol and PVP.
- Phenol:chloroform:isoamyl alcohol extraction for contaminant removal.
- RNase A treatment and isopropanol precipitation for DNA isolation.
Main Results:
- High-molecular-weight DNA obtained with A260/280 ratios of 1.78-1.98.
- Significantly higher DNA yield and band intensity compared to conventional methods (p < 0.001).
- Intact DNA with minimal RNA contamination confirmed by gel electrophoresis.
Conclusions:
- The optimized CTAB protocol offers a simple, cost-effective, and reproducible method.
- It yields high-quality genomic DNA suitable for PCR, genotyping, and sequencing.
- This protocol enhances citrus genetic research capabilities.
