Buffer Prep: Difference between revisions
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(New page: =1 M Tris-HCl Buffers= {|border="1" |+ 1 M Tris-HCl Buffers !pH !!Volume (L) !!TrisBase(g) !!HCl (ml) |- !7.0 |2 ||242.2 ||150-155 |- !7.5 |2 ||242.2 ...) |
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!4x Upper gel buffer 0.5 M Tris-Cl, pH 6.8, 0.4% SDS | !4x Upper gel buffer 0.5 M Tris-Cl, pH 6.8, 0.4% SDS | ||
|2 ||121.1 ||70-80 ||80 ml | |2 ||121.1 ||70-80 ||80 ml | ||
==10% SDS== | |||
1L: | |||
100g SDS into 1 L, heat to 68 degrees Celsius for solubility. | |||
pH ~6.6. | |||
Revision as of 13:58, 17 September 2009
1 M Tris-HCl Buffers
| pH | Volume (L) | TrisBase(g) | HCl (ml) |
|---|---|---|---|
| 7.0 | 2 | 242.2 | 150-155 |
| 7.5 | 2 | 242.2 | 120-125 |
| 8.0 | 2 | 242.2 | 80-85 |
Autoclavable.
EDTA 0.5 M (pH8.0)
0.5M, 1L: 148 g EDTA
+ ~30-40 g NaOH to adjust pH
(or 186 g EDTA-Na.2H2O + ~20 g NaOH)
Note: pH adjusted by NaOH is essential for solubility.
Autoclavable.
TAE DNA Electrophoresis Buffer(50 X)
(2 M Tris, 50 mM EDTA) 2 L 484 g Tris 114.2 ml glacial acetic acid 200 ml 0.5 M EDTA 8.0 To make 1x TAE 20 L, add 400 ml 50X buffer into 19.6 L ddH2O.
SDS-PAGE Gel Solutions
| Solution | Vol (L) | Tris (g) | HCl (ml) | 10% SDS (ml) |
|---|---|---|---|---|
| 4x Lower gel buffer 1.5 M Tris-Cl, pH 8.8, 0.4% SDS | 2 | 363.3 | 50-60 | 80 ml |
| 4x Upper gel buffer 0.5 M Tris-Cl, pH 6.8, 0.4% SDS | 2 | 121.1 | 70-80 | 80 ml
10% SDS1L: 100g SDS into 1 L, heat to 68 degrees Celsius for solubility. pH ~6.6. |
