assay development
Supplemented M9 broth (M9sup)-PDF
Materials
50 mL
500 mL
1000 mL
dH2O
43 ml
430 ml
860 ml
10 X M9 salts + glucose
5 ml
50 ml
100 ml
10% Casamino acids
1 ml
10 ml
20 ml
1% Thiamine
50 µl
0.5 ml
1 ml
1 M MgSO4
100 µl
1 ml
2 ml
1...
assay development
Making frozen permanents-PDF
Materials
Cryo vials
Overnight bacterial culture
LB (50% glycerol)
Procedure
Pipette 400 μL LB 50% glycerol and 600 μL overnight culture into a cryo vial. Vortex,...
assay development
Digestion-PDF
Overview
Standard DNA single or double digest of variable volume V using NEB restriction enzymes.
Materials
X μL DNA template
10X NEB Digestion Buffer Y (Y...
assay development
Electrocompetent cells-PDF
Materials
GYT (glycerol, yeast extract, tryptone)
• 10%(v/v) glycerol
• 0.125% (w/v) yeast extract
• 0.25% (w/v) tryptone
DI water
10% Glycerol
Special Equipment
Centrifuge
...
assay development
Chung cells-PDF
Materials
Overnight E. coli culture
50 mL Falcon tubes
LB
TSS
1.5 mL centrifuge tubes
Storage:
Freezer box
Transformation:
LB or TSS with 20 mM glucose
...
assay development
E. coli Electroporation-v1-PDF
Overview
This a protocol for preparing and handling E. coli cells for electroporation. This protocol has mostly been adapted from the Noyes Lab protocol. A...
assay development
S liquid medium-PDF
Overview
For Bcc EPS production.
Materials
1000 mL:
1 g Yeast extract
10 g Na2HPO4
3 g KH2PO4
1 g K2SO4
1 g NaCl
900 mL Water
Procedure
Dissolve...
assay development
M9 agar-PDF
Materials
500 mL
1000 mL
Bacto Agar
7.5 g
15 g
dH2O
450 ml
880 ml
10 X M9 salts + glucose
50 ml
100 ml
1 M MgSO4
1 ml
2 ml
(2 mM final)
1 M CaCl2
50 µl
0.1...
assay development
T-top (0.5% agar)-PDF
Materials
500 mL:
5 g Tryptone
5 g NaCl
500 ml dH2O
Bacto agar
1000 mL:
10 g Tryptone
10 g NaCl
1000 ml dH2O
Bacto...
assay development
10 X M9 salts
Materials
1000 mL:
700 ml dH2O
128 g Na2HPO4 • 7H2O
30 g KH2PO4
5 g NaCl
10 g NH4Cl
Procedure
Add salts to water, dissolve, adjust...
assay development
Tris-HCl-PDF
1M Tris-HCl, pH 7.5
Materials
1000 mL:
121.12 g Tris base
800 mL dH2O
HCl
Procedure
Dissolve in 800 ml dH2O, adjust pH to 7.5 with HCl, add...