How do you make a 2x CTAB buffer?
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How do you make a 2x CTAB buffer?
- Add ~200 µl of glass beads (0.5 mm).
- Vortex for 1-5 min at ≥2000 rpm until no cell clumps are visible.
- Add 3.6 µl of Proteinase K (20 mg/ml).
- Incubate at 65°C for 30-60 min.
- Add 600 µl Phenol:Chloroform:Isoamyl alcohol (25:24:1, TE-buffered to pH ≥ 7, very TOXIC!!!), mix thoroughly by vortexing several seconds.
What is CTAB solution?
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. CTAB based extraction buffers are widely used when purifying DNA from plant tissues.
What is the purpose of using CTAB during DNA extraction?
One of the most commonly used methods to extract DNA from plants uses the ionic detergent cetyltrimethylammonium bromide (CTAB) to disrupt membranes and a chloroform-isoamyl alcohol mixture that separates contaminants into the organic phase and nucleic acid into the aqueous phase.
How do you make a 10x CTAB?
Make a 10% CTAB working solution (50 mL): Combine 50 mL of 0.7 M NaCl and 2.5 g of CTAB into a 50 mL polypropylene tube (Fisher Scientific cat no. 06-443-18). Rotate slowly at 60˚C for several hours to dissolve powder completely. Store at Room Temperature for up to 6 months.
What is the pH of CTAB buffer?
pH 8.0
CTAB extraction buffer
| Reagent | Amount to add (for 10 mL) | Final concentration |
|---|---|---|
| 0.5 M EDTA (pH 8.0) | 0.4 mL | 4% |
| 1 M Tris-Cl (pH 8.0) | 1 mL | 10% |
| Polyvinylpyrrolidone (PVP) (MW 40 kDa) | 0.3 g | 3% |
| β-Mercaptoethanol | 0.02 mL | 0.2% |
How is CTAB prepared?
There is the preparation of CTAB solution. 121.1 g Tris Dissolve in about 700 ml of H2O. Bring pH down to 8.0 by adding concentrated HCl (you’ll need about 50 ml). Bring total volume to 1 L with ddH2O.
How do you make a CTAB solution?
CTAB Extraction Buffer
- Pulverize 100 mg of plant sample using a liquid nitrogen chilled mortar and pestle.
- Place the homogenate into a 60°C bath for 30 min.
- Centrifuge the homogenate for 10 minutes at 10,000 x g.
- Transfer the supernatant into a clean tube and add 5 µl of RNase (10 mg/ml in water) to the lysate.
What is CTAB made of?
The main ingredient in imitation crab is surimi, which is typically mixed with water, starch, sugar, egg whites, vegetable oil, salt and additives.
How do you make a 2% CTAB?
Fill up to 200 ml with ddH2O. Notes: CTAB is Hexadecyltrimethylammonium bromide. Dissolve it before adding NaCl, with stirring and a little warmth, if necessary.
How do I make a 1 CTAB buffer?
Is CTAB a surfactant?
Cetrimonium bromide ([(C16H33)N(CH3)3]Br; cetyltrimethylammonium bromide; hexadecyltrimethylammonium bromide; CTAB) is a quaternary ammonium surfactant. It is one of the components of the topical antiseptic cetrimide.
What is the composition for CTAB of A?
CTAB extraction buffer
| Reagent | Amount to add (for 10 mL) | Final concentration |
|---|---|---|
| Cetyltrimethyl ammonium bromide (CTAB) (10% in H2O)a | 3 mL | 3% |
| 5 M NaCl | 2.8 mL | 28% |
| 0.5 M EDTA (pH 8.0) | 0.4 mL | 4% |
| 1 M Tris-Cl (pH 8.0) | 1 mL | 10% |
Is CTAB a detergent?
CTAB is a cationic detergent, soluble in H2O and readily soluble in alcohol.
Is CTAB soluble in water?
According to the MSDS: CTAB’s solubility in water is 36.4 g/l at 20 °C (68 °F) – completely soluble. It shouldn’t require anything special.
What type of surfactant is CTAB?
Is CTAB a polymer?
Cationic surfactant (CTAB) a multipurpose additive in polymer-based drilling fluids.
