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Molecular Biology Solutions
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20x SSC (Salt sodium citrate) (1L)
175.3 g NaCl
88.2 g sodium citrate
Dissolve in ~800 ml dH2O.
Adjust pH to 7.4 with NaOH
pH should be around 7
Adjust volume to 1 liter, then dispense into smaller amounts and autoclave.
50x TAE (Tris acetate EDTA) (1L)
242 g Tris base
57.1 ml glacial acetic acid
100 ml 0.5 M EDTA (pH 8.0)
CAUTION! Glacial acetic acid is quite strong. Wear gloves and handle in hood.
10x TBE Buffer (1L)
from Maniatis, B.23
54 g Tris base
27.5 g boric acid
25 ml 0.4 M EDTA (pH 8.0)
Final concentration in 1x:
45 mM Tris-borate
45 mM boric acid
1 mM EDTA
3 M Potassium; 5 M Acetate solution for DNA purification:
Add 29.44 g of potassium acetate to 60 ml dHSUB>2O.
Then add 11.5 ml glacial acetic acid and bring to a final volume of 100 ml with dH2O.
3M Sodium Acetate
40.8 g Na-acetate-3H2O
Dissolve in ~80 ml. Adjust to pH 5.2 (or to pH 7.0 depending on what's desired), with glacial acetic acid.
Bring volume to 100 ml. Filter sterilize.
Ammonium acetate
Dissolve 0.77 g ammonium acetate in 8 ml dH2O. Adjust to 10 ml with water and filter sterilize.
1 M MgCl2 (100 ml). Autoclave.
1 M CaCl2 (100 ml). Autoclave.
1 M KCl (100 ml). Autoclave.
1 M Tris (100 ml)
12.1 g Tris base in ~80 ml dH2O.
pH to 7.0 (or 7.5 depending on what's desired) and adjust volume to 100 ml.
Autoclave.
1 M Glucose
9 g glucose in ~45 ml dH2O. Adjust volume to 50ml and filter sterilize.
SOC (500 ml)
10 g bactotryptone
2.5 g yeast extract
0.25 g NaCl
Dissolve in ~450 ml dH2O.
Add 1.25 ml 1 M KCl.
pH to 7.0 with ~0.5 ml 1 M NaOH.
Adjust volume to 500 mls. Autoclave.
Before use: For 100 ml, add 1 ml 1 M MgCl2 and 2 ml 1 M glucose.
http://research.biology.arizona.edu/mosquito/willott/procedures.html
copyright 1999-2000, Willott
Last update Sept 19, 2000
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