CTAB Protocol for Isolating DNA From Plant Tissues
To overcome the challenges presented by plant tissues, the cetyltrimethylammonium bromide (CTAB) method has become the “go-to” protocol for DNA extraction and purification from leaves and seeds. It was developed in the 1980s and has been used ever since, with various modifications for different plant species. 2-5.
CTAB Protocol for Isolating DNA from Plant Tissues
CTAB buffers are effective at removing polysaccharides and polyphenols from plant DNA preparations. CTAB (also called hexadecyltrimethylammonium bromide) is a cationic detergent that facilitates the separation of polysaccharides during purification while additives, such as polyvinylpyrrolidone, aid in inactivating polyphenols.
We use this method for extracting genome sequencing quality (i.e. unsheared) DNA that can be used for large insert libraries. It was used to extract material for the Micromonas RCC299 complete genome sequencing project, and the Micromonas RCC472 genome sequencing project.
Plant DNA Extraction using CTAB Buffer - Genetic Education
2019年2月17日 · “ Plant DNA extraction needs a CTAB DNA extraction buffer that contains ingredients that can effectively remove polysaccharides and polyphenols. Let’s see how it works and how to optimize the CTAB buffer for plant DNA extraction.”
Prepare CTAB buffer, use within 2-3 days, store capped: Add polyvinylpyrrolidone (Fisher Cat#: BP431-500) and b-mercaptoethanol (Fisher Cat#: BP176-100) and stir to dissolve right before starting extractions: 2. Weigh out 10-20 mg of silica-dried plant tissue. 3. b. washed and autoclaved sand. 4.
ue to polysaccharide production. The CTAB procedure would likely work with many gram-negative strains without modification, but gram positives would likely require the addition of lysozyme and proteinase K steps, and may also require modification of medium – eg. addition of glycine a.
What is the “modified” CTAB protocol? Characterizing ...
Cetyltrimethylammonium bromide (CTAB)–based methods are widely used to isolate DNA from plant tissues, but the unique chemical composition of secondary metabolites among plant species has necessitated optimization.