Griffitts:Stock solutions: Difference between revisions
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==Media Additives== | ==Media Additives== | ||
===1 M Calcium Chloride (75 mL)=== | ===1 M Calcium Chloride (75 mL)=== | ||
* 75 mL | * 75 mL ddH<sub>2</sub>O | ||
* 11.027 g CaCl<sub>2</sub>·2H<sub>2</sub>O | * 11.027 g CaCl<sub>2</sub>·2H<sub>2</sub>O | ||
Autoclave | Autoclave | ||
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===0.2 M IPTG=== | ===0.2 M IPTG=== | ||
* 5 mL | * 5 mL ddH<sub>2</sub>O | ||
* 0.238g isopropyl-β-D-thiogalactopyranoside (IPTG)(in freezer) | * 0.238g isopropyl-β-D-thiogalactopyranoside (IPTG)(in freezer) | ||
Sterile filter, aliquot, and store at -20°C | Sterile filter, aliquot, and store at -20°C | ||
Line 155: | Line 155: | ||
===20% SDS (75 mL)=== | ===20% SDS (75 mL)=== | ||
* 75 mL | * 75 mL ddH<sub>2</sub>O | ||
* 15 g Sodium dodecyl sulfate (SDS) | * 15 g Sodium dodecyl sulfate (SDS) | ||
Do not sterilize | Do not sterilize | ||
===5 M NaCl (75 mL)=== | ===5 M NaCl (75 mL)=== | ||
* 75 mL | * 75 mL ddH<sub>2</sub>O | ||
* 21.915 g NaCl | * 21.915 g NaCl | ||
Autoclave | Autoclave | ||
===80% Glycerol (75 mL)=== | |||
* 75 mL ddH<sub>2</sub>O | |||
* 60 g glycerol | |||
Note: use heating to mix<br> | |||
Autoclave | |||
===Orcinol spray=== | |||
* 5 mL ddH<sub>2</sub>O | |||
* 25 mL methanol (in fume hood) | |||
* 10 mL sulfuric acid (H<sub>2</sub>SO<sub>4</sub> (in fume hood—be careful!) | |||
* 200 mg orcinol |
Revision as of 10:48, 13 February 2008
Antibiotics
Ampicillin (40 mL)
- 1 g Ampicillin
- 40 mL ddH2O
- Filter sterilize
- Store in refrigerator
Chloramphenicol (40 mL)
- 0.3 g Chloramphenicol
- 40 mL 50% Ethanol
- Do not filter sterilize
- Store in refrigerator
Gentamycin (40 mL)
- 0.3 g Gentamycin
- 40 mL ddH2O
- Filter sterilize
- Store in refrigerator
Hygromycin (40 mL)
Kanamycin (40 mL)
- 0.1 g Kanamycin
- 40 mL ddH2O
- Filter sterilize
- Store in refrigerator
Neomycin (40 mL)
- 0.8 g Neomycin
- 40 mL ddH2O
- Filter sterilize
- Store in refrigerator
Spectinomycin (40 mL)
- 0.8 g Spectinomycin
- 40 mL ddH2O
- Filter sterilize
- Store in refrigerator
Streptomycin (40 mL)
- 2.0 g Streptomycin
- 40 mL ddH2O
- Filter sterilize
- Store in refrigerator
Tetracycline (40 mL)
- 0.1 g Tetracycline
- 40 mL 50% Ethanol
- Do not filter sterilize
- Store in refrigerator
Antibiotic | Abbr. | Stripe | Stock | E. coli | S. meliloti |
---|---|---|---|---|---|
Ampicillin | Ap | BLACK | 25 mg/mL | 100 μg/mL (250X) | N/A |
Chloramphenicol | Cm | BROWN | 7.5 mg/mL | 30 μg/mL (250X) | N/A |
Gentamycin | Gm | PURPLE | 7.5 mg/mL | 3 μg/mL (2500X) | 15 μg/mL (500X) |
Hygromycin | Hy | (none) | 5 mg/mL | 20 μg/mL (250X) | 20 μg/mL (250X) |
Kanamycin | Km | YELLOW | 7.5 mg/mL | 30 μg/mL (250X) | N/A |
Neomycin | Nm | GREEN | 20 mg/mL | N/A | 80 μg/mL (250X) |
Spectinomycin | Sp | ORANGE | 20 mg/mL | 80 μg/mL (250X) | 80 μg/mL (250X) |
Streptomycin | Sm | BLUE | 50 mg/mL | N/A | 200 μg/mL (250X) |
Tetracycline | Tc | RED | 2.5 mg/mL | 10 μg/mL (250X) | 10 μg/mL (250X) |
Note: These parameters are intended to be compatible with pre-made 250-mL amounts of LB agar, such that usually 1 mL of stock solution is diluted into the LB agar. For liquid LB (typically 75 mL amounts) 0.3 mL of the stock solution will usually be added. Exception: Gentamycin for E. coli.
DNA
10 mM dNTPs
- Thaw and mix (stir) 100 mM stock solutions of G, A, T, C (NEB)
- Combine in a 1.5-mL tube:
Note: Store in the -20°C freezer in the Box marked "Other Enzymes".
10X Taq buffer
Note: Store in the -20°C freezer in the Box marked "Other Enzymes".
10X Pfx buffer
Note: Store in the -20°C freezer in the Box marked "Other Enzymes".
20X Triton X-100
Media Additives
1 M Calcium Chloride (75 mL)
- 75 mL ddH2O
- 11.027 g CaCl2·2H2O
Autoclave
2% X-gal
- 10 mL N,N-dimethylformamide (DMF)(in fume hood)
- 0.2 g 5-bromo-4-chloro-3-indolyl- beta-D-galactopyranoside (X-gal)(in freezer)
Store at -20°C
0.2 M IPTG
- 5 mL ddH2O
- 0.238g isopropyl-β-D-thiogalactopyranoside (IPTG)(in freezer)
Sterile filter, aliquot, and store at -20°C
Copper sulfate
20% SDS (75 mL)
- 75 mL ddH2O
- 15 g Sodium dodecyl sulfate (SDS)
Do not sterilize
5 M NaCl (75 mL)
- 75 mL ddH2O
- 21.915 g NaCl
Autoclave
80% Glycerol (75 mL)
- 75 mL ddH2O
- 60 g glycerol
Note: use heating to mix
Autoclave
Orcinol spray
- 5 mL ddH2O
- 25 mL methanol (in fume hood)
- 10 mL sulfuric acid (H2SO4 (in fume hood—be careful!)
- 200 mg orcinol