Optimized Protocol for DNA Extraction in Three Theobroma Species
DNA extraction is a crucial step in molecular biology research, particularly for genetic and genomic analyses. These studies require a high concentration of high-quality DNA, which is often a challenge for underexplored species or when the available plant material consists of aged tissue. To address...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Bio-protocol LLC
2025-05-01
|
| Series: | Bio-Protocol |
| Online Access: | https://bio-protocol.org/en/bpdetail?id=5297&type=0 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850192890420002816 |
|---|---|
| author | Angie Riascos-España Brayan Cuastumal María Zambrano Juan Arteaga Pedro Velasquez-Vasconez |
| author_facet | Angie Riascos-España Brayan Cuastumal María Zambrano Juan Arteaga Pedro Velasquez-Vasconez |
| author_sort | Angie Riascos-España |
| collection | DOAJ |
| description | DNA extraction is a crucial step in molecular biology research, particularly for genetic and genomic analyses. These studies require a high concentration of high-quality DNA, which is often a challenge for underexplored species or when the available plant material consists of aged tissue. To address these challenges, the cetyltrimethylammonium bromide (CTAB)-based DNA extraction method has been optimized to improve efficiency and yield. The process begins with an overnight incubation of plant tissue macerated with liquid nitrogen in a solution containing a high concentration of CTAB (4%). Subsequently, the mixture undergoes two washes with chloroform: isoamyl alcohol. The nucleic acids are then precipitated using isopropanol, followed by a wash with 70% ethanol to ensure purity. Finally, the purified DNA is resuspended in ultrapure water. This optimized procedure produces high-quality DNA suitable for various downstream applications, including PCR and sequencing, even from older leaves of the three Theobroma species: T. cacao, T. bicolor, and T. grandiflorum. Additionally, this protocol significantly enhances throughput and allows for the parallel processing of a substantially larger number of samples compared to conventional techniques. |
| format | Article |
| id | doaj-art-ba588669f1a54566893c32a9951ec2bd |
| institution | OA Journals |
| issn | 2331-8325 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Bio-protocol LLC |
| record_format | Article |
| series | Bio-Protocol |
| spelling | doaj-art-ba588669f1a54566893c32a9951ec2bd2025-08-20T02:14:24ZengBio-protocol LLCBio-Protocol2331-83252025-05-0115910.21769/BioProtoc.5297Optimized Protocol for DNA Extraction in Three Theobroma SpeciesAngie Riascos-España0Brayan Cuastumal1María Zambrano2Juan Arteaga3Pedro Velasquez-Vasconez4Faculty of Exact and Natural Sciences/Biology Program, BIOGEN research group, University of Nariño, Pasto, ColombiaFaculty of Exact and Natural Sciences/Chemistry program, BIOGEN research group, University of Nariño, Pasto, ColombiaDepartment of Production and Plant Protection, BIOGEN research group, University of Nariño, Pasto, ColombiaFaculty of Exact and Natural Sciences/Chemistry program, BIOGEN research group, University of Nariño, Pasto, ColombiaDepartment of Production and Plant Protection, BIOGEN research group, University of Nariño, Pasto, ColombiaDNA extraction is a crucial step in molecular biology research, particularly for genetic and genomic analyses. These studies require a high concentration of high-quality DNA, which is often a challenge for underexplored species or when the available plant material consists of aged tissue. To address these challenges, the cetyltrimethylammonium bromide (CTAB)-based DNA extraction method has been optimized to improve efficiency and yield. The process begins with an overnight incubation of plant tissue macerated with liquid nitrogen in a solution containing a high concentration of CTAB (4%). Subsequently, the mixture undergoes two washes with chloroform: isoamyl alcohol. The nucleic acids are then precipitated using isopropanol, followed by a wash with 70% ethanol to ensure purity. Finally, the purified DNA is resuspended in ultrapure water. This optimized procedure produces high-quality DNA suitable for various downstream applications, including PCR and sequencing, even from older leaves of the three Theobroma species: T. cacao, T. bicolor, and T. grandiflorum. Additionally, this protocol significantly enhances throughput and allows for the parallel processing of a substantially larger number of samples compared to conventional techniques.https://bio-protocol.org/en/bpdetail?id=5297&type=0 |
| spellingShingle | Angie Riascos-España Brayan Cuastumal María Zambrano Juan Arteaga Pedro Velasquez-Vasconez Optimized Protocol for DNA Extraction in Three Theobroma Species Bio-Protocol |
| title | Optimized Protocol for DNA Extraction in Three Theobroma Species |
| title_full | Optimized Protocol for DNA Extraction in Three Theobroma Species |
| title_fullStr | Optimized Protocol for DNA Extraction in Three Theobroma Species |
| title_full_unstemmed | Optimized Protocol for DNA Extraction in Three Theobroma Species |
| title_short | Optimized Protocol for DNA Extraction in Three Theobroma Species |
| title_sort | optimized protocol for dna extraction in three theobroma species |
| url | https://bio-protocol.org/en/bpdetail?id=5297&type=0 |
| work_keys_str_mv | AT angieriascosespana optimizedprotocolfordnaextractioninthreetheobromaspecies AT brayancuastumal optimizedprotocolfordnaextractioninthreetheobromaspecies AT mariazambrano optimizedprotocolfordnaextractioninthreetheobromaspecies AT juanarteaga optimizedprotocolfordnaextractioninthreetheobromaspecies AT pedrovelasquezvasconez optimizedprotocolfordnaextractioninthreetheobromaspecies |