Griffitts:Stock solutions

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(Antibiotics: added a table)
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===Chloramphenicol (40 mL)===
===Chloramphenicol (40 mL)===
* 0.3 g [[Wikipedia:Chloramphenicol|Chloramphenicol]]
* 0.3 g [[Wikipedia:Chloramphenicol|Chloramphenicol]]
-
* 40 mL [[Wikipedia:Ethanol|Ethanol]]
+
* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
** Do '''not''' filter sterilize
** Do '''not''' filter sterilize
** Store in refrigerator
** Store in refrigerator
Line 28: Line 28:
** Store in refrigerator
** Store in refrigerator
===Spectinomycin (40 mL)===
===Spectinomycin (40 mL)===
-
* 20 mg [[Wikipedia:Spectinomycin|Spectinomycin]]
+
* 0.8 g [[Wikipedia:Spectinomycin|Spectinomycin]]
* 40 mL ddH<sub>2</sub>O
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
* Filter sterilize
Line 39: Line 39:
===Tetracycline (40 mL)===
===Tetracycline (40 mL)===
* 0.1 g [[Wikipedia:Tetracycline|Tetracycline]]
* 0.1 g [[Wikipedia:Tetracycline|Tetracycline]]
-
* 40 mL [[Wikipedia:Ethanol|Ethanol]]
+
* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
** Do '''not''' filter sterilize
** Do '''not''' filter sterilize
** Store in refrigerator
** Store in refrigerator
Line 73: Line 73:
! style="background:#9400D3;" | <font color=white>'''PURPLE'''</font>
! style="background:#9400D3;" | <font color=white>'''PURPLE'''</font>
| 7.5 mg/mL
| 7.5 mg/mL
-
| 3 μg/mL (250X)
+
| 3 μg/mL (2500X)
| 15 μg/mL (500X)
| 15 μg/mL (500X)
|-
|-

Revision as of 18:21, 4 February 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)

  • 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)




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 (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:
    • 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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