CTAB Lysis Buffer
CTAB Extraction Buffer
Components: CTAB (cetyltrimethylammonium bromide), Tris-HCl, EDTA, NaCl, β-mercaptoethanol, and PVP.
- CTAB is a detergent that disrupts cell membranes and forms complexes with polysaccharides, helping to separate them from D.
- Tris-HCl buffers the pH to stabilize DNA.
- EDTA chelates divalent cations (like Mg²⁺), which inactivates DNases.
- NaCl aids in removing polysaccharides and proteins.
- β-mercaptoethanol reduces disulfide bonds, preventing oxidation of phenolic compounds.
- PVP (polyvinylpyrrolidone) binds polyphenols that might otherwise co-precipitate with DNA.
| Ingredient | Concentration |
| CTAB | 1% |
| Tris | 100 mM |
| EDTA | 20 mM |
| NaCl | 1 M |
| PVP | 1% |
Preparation
CTAB Lysis Buffer
Step 1.
Use the 10% stock CTAB solution.
10% CTAB STOCK
5
mL
Step 2.
Prepare ingredients.
Tris Base
0.6057
g
(100 mM)
Step 3.
Prepare ingredients.
EDTA (Ethylenediaminetetraacetic acid)
0.2922
g
(20 mM)
Step 4.
Prepare ingredients.
Sodium Chloride
2.922
g
(1 M)
Step 5.
Prepare ingredients.
2-Mercaptoethanol
100
μL
Step 6.
Prepare ingredients.
Polyviny|pyrrolidone (PVP)
0.5
g
(1 % w/v)
Step 7.
Bring final volume to 50 mL with water.