Griffitts:Stock solutions

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m (10 mM dNTPs)
(Antibiotics: added a table)
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** Do '''not''' filter sterilize
** Do '''not''' filter sterilize
** Store in refrigerator
** Store in refrigerator
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{| border="1" cellpadding="5" cellspacing="0" class="wikitable sortable"
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|+'''Antibiotic Specifications'''
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|-
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! width="120" style="background:#efefef;" | Antibiotic
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! width="30" style="background:#efefef;" | Abbr.
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! width="90" style="background:#efefef;" | Stripe
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! width="90" style="background:#efefef;" | Stock
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! width="120" style="background:#efefef;" | ''E. coli''
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! width="120" style="background:#efefef;" | ''S. meliloti''
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|-
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| Ampicillin
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| Ap
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! style="background:#000000;" | <font color=white>'''BLACK'''</font>
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| 25 mg/mL
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| 100 μg/mL (250X)
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| N/A
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|-
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| Chloramphenicol
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| Cm
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! style="background:#8B4513;" | <font color=white>'''BROWN'''</font>
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| 7.5 mg/mL
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| 30 μg/mL (250X)
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| N/A
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|-
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| Gentamycin
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| Gm
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! style="background:#9400D3;" | <font color=white>'''PURPLE'''</font>
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| 7.5 mg/mL
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| 3 μg/mL (250X)
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| 15 μg/mL (500X)
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|-
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| Hygromycin
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| Hy
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| (none)
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| 5 mg/mL
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| 20 μg/mL (250X)
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| 20 μg/mL (250X)
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|-
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| Kanamycin
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| Km
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! style="background:#FFD700;" | <font color=white>'''YELLOW'''</font>
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| 7.5 mg/mL
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| 30 μg/mL (250X)
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| N/A
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|-
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| Neomycin
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| Nm
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! style="background:#008000;" | <font color=white>'''GREEN'''</font>
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| 20 mg/mL
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| N/A
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| 80 μg/mL (250X)
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|-
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| Spectinomycin
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| Sp
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! style="background:#FF8C00;" | <font color=white>'''ORANGE'''</font>
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| 20 mg/mL
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| 80 μg/mL (250X)
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| 80 μg/mL (250X)
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|-
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| Streptomycin
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| Sm
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! style="background:#0000FF;" | <font color=white>'''BLUE'''</font>
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| 50 mg/mL
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| N/A
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| 200 μg/mL (250X)
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|-
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| Tetracycline
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| Tc
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! style="background:#FF0000;" | <font color=white>'''RED'''</font>
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| 2.5 mg/mL
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| 10 μg/mL (250X)
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| 10 μg/mL (250X)
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|}
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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 16:49, 28 January 2008

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Contents

Antibiotics

Ampicillin (40 mL)

  • 1 g Ampicillin
  • 40 mL ddH2O
  • Filter sterilize
    • Store in refrigerator

Chloramphenicol (40 mL)

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)




Antibiotic Specifications
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:
    • 60 μl H2O
    • 10 μl G
    • 10 μl A
    • 10 μl T
    • 10 μl C

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

X-gal

IPTG

Copper sulfate

SDS

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