Solutions for Bacteria
Tetracycline (7.5 mg/ml stock)
Tetracycline 375 mg
100% EtOH 25 ml
fill up to 50 ml with ddH2O
Ampicillin (100 mg/ml stock)
Ampicillin 5 g
fill up to 50 ml with ddH2O
LB media
Bacto tryptone 10 g
Yeast extract 5 g
NaCl 5 g
ddH2O up to 1 L
autoclave;
add antibiotic when cooled enough to touch comfortably (~50 degrees)
15 mg/L ampicillin: add 150 ml of 100 mg/ml stock
7.5 mg/L tetracycline: add 1 ml of 7.5 mg/ml stock
for Amp only LB media (50 mg/L ampicillin): add 500 ml of 100 mg/ml stock
Amp/Tet plates; Amp plates
Bacto tryptone 10 g
Yeast extract 5 g
NaCl 5 g
Difco Agar 15 g
ddH2O up to 1 L
autoclave;
add antibiotic when cooled enough to touch comfortably (~50 degrees)
15 mg/L ampicillin: add 150 ml of 100 mg/ml stock
7.5 mg/L tetracycline: add 1 ml of 7.5 mg/ml stock
for Amp only plates (50 mg/L ampicillin): add 500 ml of 100 mg/ml stock
· pour into Petri dishes; once solidified, wrap plates with foil and store at 4 degrees.
· dry plates in 42 degree incubator before using
SOC medium
0.5% yeast extract 5 g
2% bacto tryptone 20 g
10 mM NaCl 0.58 g
2.5 mM KCl 25 ml 100 mM KCl
(0.37 g KCl in 50 ml H2O)
10 mM MgCl2 2 g
10mM MgSO4 2.46 g
20 mM Glucose 3.6 g
fill up to 1 L with ddH2O; autoclave
Solution #1
[final]
0.5 M EDTA, pH 8.0 20 ml 10 mM
1 M Tris, pH 7.5 25 ml 25 mM
glucose 9 g 50 mM
bring up volume to 1 L with ddH2O
autoclave
Solution #2
[final]
ddH2O 88 ml
10% SDS 10 ml 1%
10 M NaOH 2 ml 200 mM
Solution #3
[final]
Potassium Acetate 250 g 2.5 M
Glacial Acetic Acid 150 ml
pH should be ~4.7
bring up volume to 1 L with ddH2O
TE/RNase
[final]
1 M Tris (pH 7.5) 500 ml 10 mM
0.5 M EDTA (pH 8) 100 ml 1 mM
bring up volume to 50 ml ddH2O
* to destroy contaminating RNA, add 0.5 ml of 20 mg/ml RNase A to 50 ml of TE (final 200 mg/ml)
Updated 3/11/10 by RK