High-Yield-Plasmid-Medium
Low Yield LB medium (g/L):
[edit]tryptone, 10; yeast extract, 5.0; NaCl, 10.0.
High Yield Medium (g/L):
[edit]Tryptone, 10.0; yeast extract, 10 or 15; NaCl, 10.0--> 930 ml H2O;
or 25 g LB mix powder + 10 g yeast extract --->930 ml H2O;
Trace metal solution, 1 ml/L;
50 X 2G, 20 ml;
20 X P, 50 ml;
Ampicillin, 100 mg/L, i.e., 1 ml of 1000X Amp or Carbenicillin;
or Kanamycin, 50 mg/L, i.e., 1 ml of 1000 X Kan;
Antiform, one tiny drop (don't add more than 5 ul).
50 X 2G (g/L)
Glucose, 200;
Glycerol, 100;
Water to 1 L;
Filter sterilize.
(1X: glycerol, 2.0; glucose, 4 or 5)
20 X P (g/L)
K2HPO4, 10;
KH2PO4, 125;
(NH4)2HPO4, 40;
MgSO4, 20;
Water to 1L;
Autoclave.
(1 X: K2HPO4, 0.5; KH2PO4, 2.5; (NH4)2HPO4, 2.0; MgSO4, 1.0)
Trace metal solution (g/L):
FeCl3·6H2O, 0.162;
ZnCl2·4H2O, 0.0144;
CoCl2·6H2O, 0.12;
Na2MoO4·2H2O, 0.012;
CaCl2·2H2O, 0.006;
CuSO4·5H2O, 1.9;
H3BO3, 0.5;
HCl (37%, w/v), 37 ml/L
Filter sterilize
TB +GP
[edit]12 g tryptone
24 g yeast extract,
4 ml glycerol, water to 900 ml, autoclave
Then add 20 mL filter sterilized 1 M KH2PO4 and 80 mL filter sterilized 1 M K2HPO4
2XYT + GP
[edit]10 g yeast extract
16 g tryptone (or peptone)
5g NaCl
4 ml glycerol, water to 900 ml, autoclave
Then add 20 mL filter sterilized 1 M KH2PO4 and 80 mL filter sterilized 1 M K2HPO4
LB + GP
[edit]5g yeast extract
10 g tryptone (or peptone)
10 g NaCl
4 ml glycerol, water to 900 ml, autoclave
Then add 20 mL filter sterilized 1 M KH2PO4 and 80 mL filter sterilized 1 M K2HPO4
For Plasmid Production
[edit]1 Liter
K2HPO4, 12.54 g;
KH2PO4, 2.31 g;
D-Glucose, 4.0g;
NH4Cl, 1.0g;
Yeast extract, 12.0g;
NaCl, 5.0g;
MgSO4, 0.24g;