Team:Goettingen/Project/Materials
From 2012.igem.org
(Difference between revisions)
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===== Genetical Characteristics ===== | ===== Genetical Characteristics ===== | ||
* Antibiotic resistance: none | * Antibiotic resistance: none | ||
+ | ===== Reference ===== | ||
+ | * <html><a href="http://www.invitrogen.com/1/3/dh5a-competent-cells">Invitrogen</a></html> | ||
<br> | <br> | ||
==== BL21DE3 ==== | ==== BL21DE3 ==== | ||
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* Antibiotic resistance: none! | * Antibiotic resistance: none! | ||
===== Reference ===== | ===== Reference ===== | ||
- | * <html><a href="http://www.merckmillipore.com/germany/life-science-research/bl21de3-glycerol-stock/EMD_BIO-69387/p_52yb.s1OQ1MAAAEj7Bt9.zLX">Merck</a></html> | + | * <html><a href="http://www.merckmillipore.com/germany/life-science-research/bl21de3-glycerol-stock/EMD_BIO-69387/p_52yb.s1OQ1MAAAEj7Bt9.zLX">Merck Millipore</a></html> |
<br> | <br> | ||
==== XL1 blue ==== | ==== XL1 blue ==== | ||
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===== Genetical Characteristics ===== | ===== Genetical Characteristics ===== | ||
* Antibiotic resistance: none! | * Antibiotic resistance: none! | ||
+ | ===== Reference ===== | ||
+ | <html><a href="http://www.merckmillipore.com/life-science-research/competent-cells-spotlight-main/c_CzSb.s1O1SAAAAEhSc8sgj.x?back=true&cid=bios-x-goog-1UKEN1277>Merck Millipore</a></html> | ||
<br> | <br> | ||
==== MG1655 ==== | ==== MG1655 ==== |
Revision as of 16:11, 26 September 2012
Deutsch / English |
Contents |
Recipes
Media
Recipes for liquid and solid media for bacterial cultivation. Stir until dissolved; sterilize by autoclaving 121 °C. Do not forget a magnetic stirrer.
LB-broth
- 1 % (w/v) bacto tryptone
- 0.5 % (w/v) bacto yeast extract
- 1 % (w/v) NaCl
- if required: ad 1 % (w/v) agar
M9 Swimming Agar
- 1.25 % (v/v) glycerol
- 20 % (v/v) 5x M9 salt stock solution
- 0.1 % (v/v) of CaCl2 x 2 H2O stock solution (20 mg/mL)
- 0.1 % (v/v) MgSO4 stock solution (0.12 g/mL)
- 0.3 % (w/v) of agar
Tryptone swimming agar
- 1 % (w/v) bacto tryptone
- 0.5 % (w/v) NaCl
- 0.3 % (w/v) agar
Antibiotics
- Stock solutions and concentrations of antibiotics.
Reagent | Abbreviation | Stock Conc. [mg/mL] | Working Conc. [µg/mL] | Dilution | Solvent | Notes |
---|---|---|---|---|---|---|
Ampicillin | Amp | 100 | 100 | 1,000x | ddH2O | Culture plates with Amp can be stored at 4 °C for about 2 weeks. Stock solutions can be stored at 4 °C for 2 weeks but can last as long as 4-6 months when stored at -20 °C. |
Chloramphenicol | Cm | 20 | 20 | 1,000x | EtOH | |
Kanamycin | Kan | 50 | 50 | 1,000x | ddH2O | Plates may only last a few weeks at 4 °C. |
Supplements
- Aminoacid mix (containing all aminoacids except tryptophane): stocksolution 0.2 % (w/v), varying amounts used in swimming assays, view methods and notebook of group 1
- Caffeine: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
- D-Aspartate: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
- 2-Ethyl-1-Hexanole: stocksolution 5 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
- Geraniol: stocksolution 5 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
- L-Aspartate-4-Benzylester: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
- L-aspartate: stocksolution 10 mM, varying amounts used in swimming assays, view methods and notebook of group 1
- Leucin: stocksolution 30.49 mM, add to M9-agar: 4 mL (final concentration 0.305 µM)
- Methionin: stocksolution 26.8 mM, add to M9-agar: 4 mL (final concentration 0.268 µM)
- Sodium Cyclamate: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
- Tryptone: stocksolution 0.5 % (w/v), varying amounts used in swimming assays, view methods and notebook of group 1
- Vanillin: stocksolution 10 mM, 100 µL applied to whatmanpaper in swimming assays, view methods
Buffer and Solution
Recipes for stocks of buffers and solutions.
5x M9 salts stock solution
Amount | Substance |
---|---|
64 g | Na2HPO4 × 7 H2O |
15 g | KH2PO4 |
2.5 g | NaCl |
5.0 g | NH4Cl |
ad 1000 ml | ddH2O |
- Stir until dissolved
CaCl2 Buffer for competent cells
Amount | Substance | Notes |
---|---|---|
7.351 g | CaCl2 × 7 H2O | Alternative 0.1 M CaCl2 |
15% | Glycerol | |
ad 500 ml | ddH2O |
- Stir until dissolved; sterilize by sterile filtration.
50x TAE for agarose gels
Amount | Substance |
---|---|
242 g | Tris base |
57.1 mL | Glacial acetic acid |
18.6 g | EDTA |
ad 1000 mL | ddH2O |
- Stir until dissolved. Working concentration is 1x (40 mL of 50x stock ad 2 L ddH2O).
E. coli
- E. coli strain list - List of the working E. coli strains including genotypes, genetic markers, and alleles.
Non-mutated strains
These strains may not contain any plasmids or resistances, because they should be used for transformation or other experiments.
DH10B
Genotype
- F- endA1 recA1 galE15 galK16 nupG rpsL ΔlacX74 Φ80lacZΔM15 araD139 Δ(ara,leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) λ-
Genetical Characteristics
- Antibiotic resistance: none
Reference
DH5α
Genotype
- F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF)U169, hsdR17(rK- mK+), λ–
Genetical Characteristics
- Antibiotic resistance: none
Reference
BL21DE3
Genotype
- E. coli B F- dcm ompT hsdS(rB- mB-) gal [malB+]K-12(λS)
Genetical Characteristics
- Antibiotic resistance: none!
Reference
XL1 blue
Genotype
- endA1 gyrA96(nalR) thi-1 recA1 relA1 lac glnV44 F'[ ::Tn10 proAB+ lacIq Δ(lacZ)M15] hsdR17(rK- mK+)
Genetical Characteristics
- Antibiotic resistance: none!
Reference
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