培養基與各種試劑的配製

100mg/ml Amp Agar plate
15 g agar 溶於 1000 ml YT medium
121°C ，高壓滅菌 40 分鐘，冷卻約 55°C ，加入 1 ml ，100mg/ml
ampicillin，混合均勻，再將溶液倒於 (90x15mm) 培養皿，靜置於平衡台，待凝固，方可使用。
---------------------------------------------
Lysis Buffer
urea 8M
NaH 2 PO 4 100mM
Tris⋅Cl 10mM
以 10N NaOH 調至 pH 8.0
-----------------
CAPS transfer buffer, 500ml
CAPS 10mM
Methanol 50ml
以10N NaOH 調至 pH 11.0
---------------
Coomassie Staining Solution, 500ml
Fixing solution 500ml
Coomassie brilliant blue R-250 0.25g
--------------
Destaining Solution, 500ml
methanol 25ml
acetate acid 35 ml
H2O to 500ml
----------------
6x DNA loading dye, 15ml
100% glycerol 4.5ml
bromophenol blue 0.0375g
xylene cyanol 0.0375g
H2O to 15ml
-----------------
Fixing Solution I , 200ml
methanol 80ml
acetate acid 20ml
H2O to 200ml
---------------
Fixing Solution II , 200ml
ethenol 42ml
acetate acid 20ml
H2O to 200ml
-----------------
5x M9 medium, 500ml
Na2HPO4 15g
KH2PO4 7.5g
NaCl 1.25g
NH4Cl 2.5g
1M CaCl2 68ml
H2O to 500ml
----------------
M9 medium Need, 2 ml
1 M MgSO4 0.5ml
100% glycerol 1ml
5% vitamine B1 5ml
40mg/ml L-amino acid 0.5ml
加水至 2 ml
------------
M9 plate, 500 ml
5x M9 medium 100ml
agar 7.5g
H2O to 500ml
121°C ，高壓滅菌 40 分鐘，冷卻約 55°C ，加入 2 ml M9 medium Need，混合均勻，再將溶液倒於 (90×15mm) 培養皿，靜置於平衡台，待凝固，方可使用。
------------
10x Phosphate Buffered Saline (PBS), 1000 ml
NaCl 80g
KCl 2g
Na2HPO4⋅7H2O 15.4g
KH2PO4 2g
H2O to 1000ml
調至 pH 7.3， autoclave。
--------------------------
10x Tricine-SDS/Electrophoresis buffer, 1000 ml (pH 8.3)
Tris⋅base 120g
tricine 179g
SDS 10g
加水至1000 ml
------------------
2x SDS sample buffer, dH 2 O to 10 ml
3M Tris⋅HCl, pH 8.45 3.0ml
bromphenol blue 0.2g
SDS 0.8gm
80% glycerol 3.0ml
0.1% Phenol Red 0.2g
------------------------
Solution I, 100 ml
1M glucose 5ml
1M Tris⋅HCl, pH 8.0 2.5ml
0.5M EDTA, pH 8.0 2ml
混合後，用 0.2µm 濾膜過濾。
---------------------
Solution II
0.2 N NaOH
1% SDS
--------------------
Solution III, 100 ml
5 M potassium acetate 60ml
99.8% acetic acid 11.5ml
H2O to 100 ml
-----------------------
20x SSC, pH 7.0, 1000 ml
NaCl 175g
Na3⋅Citrate 88g
H2O to 1000ml
Autoclave
------------------------
10x TBE buffer, 1000 ml
Tris⋅Base 108g
boric acid 5.5g
0.5M EDTA, pH 8.0 40ml
----------------------
1x T.E. buffer, pH 8.0, 100ml
1M Tris⋅HCl, pH 8.0 1ml
0.5M EDTA, pH 8.0 0.2ml
H2O to 100ml
Autoclave
------------------
Transfer buffer, pH 6.8, 1000 ml
Tris⋅base 5.81g
glycine 2.93g
SDS 0.375g
methanol 200ml
H2O 1000ml
---------------------
Wash buffer
urea 8M
Na 2 HPO 4 50mM
NaCl 100mM
pipes 20mM
pH值 調至 pH 6.34
----------------------
YT medium, 1000 ml
trypton 10g
yeast extract 5g
NaCl 5g
以 NaOH 調至 pH 7.4， 121°C，高壓滅菌 40 分鐘。
--------------------------
YT plate
15 g agar 溶於 1000 ml YT medium
121°C ，高壓滅菌 40 分鐘，冷卻約 55°C ，將溶液倒於 (90x15mm) 培養皿，靜置於平衡台，待凝固，方可使用。
--------------------------
Tyrode solution, pH 7.4
NaCl 136.9 mM
KCl 2.7 mM
MgCl 2 2.1 mM
NaH 2 PO 4 0.4 mM
NaHCO 3 11.9 mM