Griffitts:Stock solutions: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
m (→10 mM dNTPs) |
(→Antibiotics: added a table) |
||
Line 42: | Line 42: | ||
** Do '''not''' filter sterilize | ** Do '''not''' filter sterilize | ||
** Store in refrigerator | ** Store in refrigerator | ||
<br> | |||
<br> | |||
<br> | |||
{| border="1" cellpadding="5" cellspacing="0" class="wikitable sortable" | |||
|+'''Antibiotic Specifications''' | |||
|- | |||
! width="120" style="background:#efefef;" | Antibiotic | |||
! width="30" style="background:#efefef;" | Abbr. | |||
! width="90" style="background:#efefef;" | Stripe | |||
! width="90" style="background:#efefef;" | Stock | |||
! width="120" style="background:#efefef;" | ''E. coli'' | |||
! width="120" style="background:#efefef;" | ''S. meliloti'' | |||
|- | |||
| Ampicillin | |||
| Ap | |||
! style="background:#000000;" | <font color=white>'''BLACK'''</font> | |||
| 25 mg/mL | |||
| 100 μg/mL (250X) | |||
| N/A | |||
|- | |||
| Chloramphenicol | |||
| Cm | |||
! style="background:#8B4513;" | <font color=white>'''BROWN'''</font> | |||
| 7.5 mg/mL | |||
| 30 μg/mL (250X) | |||
| N/A | |||
|- | |||
| Gentamycin | |||
| Gm | |||
! style="background:#9400D3;" | <font color=white>'''PURPLE'''</font> | |||
| 7.5 mg/mL | |||
| 3 μg/mL (250X) | |||
| 15 μg/mL (500X) | |||
|- | |||
| Hygromycin | |||
| Hy | |||
| (none) | |||
| 5 mg/mL | |||
| 20 μg/mL (250X) | |||
| 20 μg/mL (250X) | |||
|- | |||
| Kanamycin | |||
| Km | |||
! style="background:#FFD700;" | <font color=white>'''YELLOW'''</font> | |||
| 7.5 mg/mL | |||
| 30 μg/mL (250X) | |||
| N/A | |||
|- | |||
| Neomycin | |||
| Nm | |||
! style="background:#008000;" | <font color=white>'''GREEN'''</font> | |||
| 20 mg/mL | |||
| N/A | |||
| 80 μg/mL (250X) | |||
|- | |||
| Spectinomycin | |||
| Sp | |||
! style="background:#FF8C00;" | <font color=white>'''ORANGE'''</font> | |||
| 20 mg/mL | |||
| 80 μg/mL (250X) | |||
| 80 μg/mL (250X) | |||
|- | |||
| Streptomycin | |||
| Sm | |||
! style="background:#0000FF;" | <font color=white>'''BLUE'''</font> | |||
| 50 mg/mL | |||
| N/A | |||
| 200 μg/mL (250X) | |||
|- | |||
| Tetracycline | |||
| Tc | |||
! style="background:#FF0000;" | <font color=white>'''RED'''</font> | |||
| 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 [[Griffitts:Common media#LB Broth (1 L)|LB]] agar, such that usually 1 mL of stock solution is diluted into the [[Griffitts:Common media#LB Broth (1 L)|LB]] agar. For liquid [[Griffitts:Common media#LB Broth (1 L)|LB]] (typically 75 mL amounts) 0.3 mL of the stock solution will usually be added. Exception: Gentamycin for ''E. coli''. | |||
==DNA== | ==DNA== |
Revision as of 13:49, 28 January 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 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)
- 20 mg 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 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 (250X) | 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:
10X Taq buffer
10X Pfx buffer
20X Triton X-100
Media Additives
1 M Calcium Chloride (75 mL)
- 75 mL dH2O
- 11.027 g CaCl2·2H2O
Autoclave