VARIOUS PLATE AND PLATE SOLUTION RECIPES
- Try to use the same autoclave for an entire experiment to decrease variance.
- For all plates, cool while stirring until the flask can be touched comfortably to the inside of your arm.
Do not over-cool as the melted agar will start to gel.
- Pour plates about 30mls for 100mm x 15 mm plates and 90mls for 150mm x 15mm plates
SM Agar (1 Liter)
G418- SM Plates (1 Liter)
For .3µl/ml
G418
300µl
Oxoid Peptone
10 grams
Oxoid Yeast Extract
1 gram
Glucose
10 grams
KH2PO4
1.9 grams
K2HPO4
1.3 grams
SM/5 Agar (1 Liter)
MgSO4anhydrous
.49 grams
Oxoid Peptone
2 grams
Agar
17 grams
Oxoid Yeast Extract
0.2 grams
Glucose
2 grams
KH2PO4
1.9 grams
K2HPO4
1.3 grams
** SM Broth is without Agar
There is an alternative method:
Oxoid Peptone
10 grams
For .5µl/ml
G418
500µl
Oxoid Yeast Extract
1 gram
MgSO4anhydrous
.2 grams
KH2PO4
1.9 grams
Agar
17 grams
K2HPO4
1.3 grams
MgSO4anhydrous
.49 grams
SM/10 Agar (1 Liter)
Agar
dH2O
17 grams
750ml
Oxoid Peptone
1 gram
Oxoid Yeast Extract
.1 gram
Glucose
1 gram
KH2PO4
1.9 grams
K2HPO4
1.3 grams
MgSO4anhydrous
.1 grams
Agar
17 grams
Glucose
10g
dH2O
250ml
Autoclave the glucose in a separate flask from
the other ingredients (to avoid caramelizing the
sugars). Pour the glucose solution in when the
bottle/flask is cool enough to hold your hand
against it. Add MgSO4 after autoclaving to avoid
precipitation:
100x MgSO4stock solution
10ml
100MgSO4 stock solution
MgSO4
25 grams
dH2O
250ml
LB-Broth Miller (1Liter)
Tryptone
10 grams
Oxoid Yeast Extract
5 grams
NaCl
10 grams
Final PH
7.0 ± 0.2
SM/20 Agar (1 Liter)
Hay Plates (1 Liter) – see Hay Plate Protocol
before using this recipe
Oxoid Peptone
0.5 grams
Oxoid Yeast Extract
0.05 grams
KH2PO4
1.5 grams
Glucose
0.5 grams
Na2HPO4.7HsO
.62 grams
KH2PO4
1.9 grams
Agar
15 grams
K2HPO4
1.3 grams
MgSO4anhydrous
.05 grams
Agar
17 grams
HL5 Medium (1 Liter)
Oxoid Peptone
14 grams
Oxoid Yeast Extract
7 grams
KH2PO4
0.5 grams
Na2HPO4
0.5 grams
Glucose
31.5 grams
KK2 buffer (1 liter)
KH2PO4
2.25 grams
K2HPO4
.67 grams
TBE (1X) – 20 Liters
Tris Base
216 grams
Boric Acid
110 grams
Na2EDTA
18.6 grams
ddH2O
20 Liters
LP Agar (1 Liter)
Lactose
1 gram
Bactopeoptone
1 gram
Agar
15 grams
SS ( 1 Liter)
50x Starving Buffer (100 mls)
Na2HPO4 dibasic
1.78 grams
KH2PO4monobasic
9.9 grams
**Water Agar Plates are 1% for example for
300ml add 3 grams of Agar
Starving Plates (1 Liter)
NaCl
0.6 grams
KCL
0.75 grams
CaCl2
0.3 grams
Lactose Plates (1 Liter) – low nutrient plates for
robust strains
Lactose
1 gram
Bacto Peptone
1 gram
50x Starving Buffer
20 mls
KH2PO4
2.2 grams
H2O
980 mls
Na2HPO4.2H2O
1.25 grams
Agar
20 grams
Agar
15 grams
Adjust pH to 6.5
Spore Buffer
EDTA
20mM
NP-40
0.1%
KK2
1ml
Freezing Medium – in HL5
Sterile horse or calf
serum
DMSO
50%
7.5%
PSV (500mls)
Streptomycin
25 grams
Penicillin
5 grams
10,000x of Vitamin B12
(6mg/ml)
5 mls
10,000x of Folic Acid
(2mg/ml)
5 mls
Filter, and sterilize at 40C