Griffitts:Stock solutions

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* 40 mL ddH<sub>2</sub>O
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
* Filter sterilize
-
** Store in refrigerator
+
NOTE: Store at 4°C
===Chloramphenicol (40 mL)===
===Chloramphenicol (40 mL)===
-
* 0.3 g [[Wikipedia:Chloramphenicol|Chloramphenicol]]
+
* 300 mg [[Wikipedia:Chloramphenicol|Chloramphenicol]]
* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
-
** Do '''not''' filter sterilize
+
NOTE: Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter<br>
-
** Store in refrigerator
+
NOTE: Store at 4°C
===Erythromycin (20 mL)===
===Erythromycin (20 mL)===
-
* 1.0 g [[Wikipedia:Erythromycin|Erythromycin]]
+
* 1 g [[Wikipedia:Erythromycin|Erythromycin]]
* 20 mL 100% ethanol (in fume hood)
* 20 mL 100% ethanol (in fume hood)
* Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter
* Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter
-
** Store in the &ndash;20°C Freezer
+
NOTE: Store at &ndash;20°C
-
 
+
===Gentamicin (40 mL)===
===Gentamicin (40 mL)===
-
* 0.3 g [[Wikipedia:Gentamicin|Gentamicin]]
+
* 300 mg [[Wikipedia:Gentamicin|Gentamicin]]
* 40 mL ddH<sub>2</sub>O
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
* Filter sterilize
-
** Store in refrigerator
+
NOTE: Store at 4°C
-
 
+
===Hygromycin (40 mL)===
===Hygromycin (40 mL)===
 +
* 200 mg [[Wikipedia:Hygromycin|Hygromycin]]
 +
* 40 mL ddH<sub>2</sub>O
 +
* Filter sterilize
 +
NOTE: Store at 4°C
===Kanamycin (40 mL)===
===Kanamycin (40 mL)===
-
* 0.3 g [[Wikipedia:Kanamycin|Kanamycin]]
+
* 300 mg [[Wikipedia:Kanamycin|Kanamycin]]
* 40 mL ddH<sub>2</sub>O
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
* Filter sterilize
-
** Store in refrigerator
+
NOTE: Store at 4°C
-
 
+
===Nalidixic acid (30 mL)===
===Nalidixic acid (30 mL)===
-
* 0.9 g [[Wikipedia:Nalidixic Acid|Nalidixic Acid]]
+
* 900 mg [[Wikipedia:Nalidixic Acid|Nalidixic Acid]]
-
* 0.36 g NaOH
+
* 360 mg NaOH
* 30 mL ddH<sub>2</sub>O
* 30 mL ddH<sub>2</sub>O
* Filter sterilize
* Filter sterilize
-
** Store in refrigerator
+
NOTE: Store at 4°C
-
 
+
===Neomycin (40 mL)===
===Neomycin (40 mL)===
-
* 1.0 g [[Wikipedia:Neomycin|Neomycin]]
+
* 1 g [[Wikipedia:Neomycin|Neomycin]]
* 40 mL ddH<sub>2</sub>O
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
* Filter sterilize
-
** Store in refrigerator
+
NOTE: Store at 4°C
-
 
+
===Rifampicin (20 mL)===
===Rifampicin (20 mL)===
-
* 1.0 g [[Wikipedia:Rifampicin|Rifampicin]]
+
* 1 g [[Wikipedia:Rifampicin|Rifampicin]]
* 20 mL N,N-dimethylformamide (DMF; in fume hood)
* 20 mL N,N-dimethylformamide (DMF; in fume hood)
-
* Do '''not''' filter sterilize&mdash;the DMF will dissolve the filter
+
NOTE: Do '''not''' filter sterilize&mdash;the DMF will dissolve the filter<br>
-
** Store in the &ndash;20°C Freezer
+
NOTE: Store at &ndash;20°C
-
 
+
===Spectinomycin (40 mL)===
===Spectinomycin (40 mL)===
-
* 0.8 g [[Wikipedia:Spectinomycin|Spectinomycin]]
+
* 800 mg [[Wikipedia:Spectinomycin|Spectinomycin]]
* 40 mL ddH<sub>2</sub>O
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
* Filter sterilize
-
** Store in refrigerator
+
NOTE: Store at 4°C
-
 
+
===Streptomycin (40 mL)===
===Streptomycin (40 mL)===
-
* 2.0 g [[Wikipedia:Streptomycin|Streptomycin]]
+
* 2 g [[Wikipedia:Streptomycin|Streptomycin]]
* 40 mL ddH<sub>2</sub>O
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
* Filter sterilize
-
** Store in refrigerator
+
NOTE: Store at 4°C
===Tetracycline (40 mL)===
===Tetracycline (40 mL)===
-
* 0.1 g [[Wikipedia:Tetracycline|Tetracycline]]
+
* 100 mg [[Wikipedia:Tetracycline|Tetracycline]]
* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
-
** Do '''not''' filter sterilize
+
NOTE: Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter<br>
-
** Store in refrigerator
+
NOTE: Store at 4°C
==Antibiotics Table==
==Antibiotics Table==

Revision as of 12:40, 17 March 2010

Contents

Antibiotics

Ampicillin (40 mL)

NOTE: Store at 4°C

Chloramphenicol (40 mL)

NOTE: Do not filter sterilize—the ethanol may dissolve the filter
NOTE: Store at 4°C

Erythromycin (20 mL)

  • 1 g Erythromycin
  • 20 mL 100% ethanol (in fume hood)
  • Do not filter sterilize—the ethanol may dissolve the filter

NOTE: Store at –20°C

Gentamicin (40 mL)

NOTE: Store at 4°C

Hygromycin (40 mL)

NOTE: Store at 4°C

Kanamycin (40 mL)

  • 300 mg Kanamycin
  • 40 mL ddH2O
  • Filter sterilize

NOTE: Store at 4°C

Nalidixic acid (30 mL)

NOTE: Store at 4°C

Neomycin (40 mL)

  • 1 g Neomycin
  • 40 mL ddH2O
  • Filter sterilize

NOTE: Store at 4°C

Rifampicin (20 mL)

  • 1 g Rifampicin
  • 20 mL N,N-dimethylformamide (DMF; in fume hood)

NOTE: Do not filter sterilize—the DMF will dissolve the filter
NOTE: Store at –20°C

Spectinomycin (40 mL)

NOTE: Store at 4°C

Streptomycin (40 mL)

NOTE: Store at 4°C

Tetracycline (40 mL)

NOTE: Do not filter sterilize—the ethanol may dissolve the filter
NOTE: Store at 4°C

Antibiotics Table

Antibiotic Specifications
Antibiotic Abbr. Stripe Stock E. coli S. meliloti Solvent Storage
Ampicillin Ap BLACK 25 mg/mL 100 μg/mL (250X) N/A ddH2O 4°C
Chloramphenicol Cm BROWN 7.5 mg/mL 30 μg/mL (250X) N/A EtOH 4°C
Erythromycin Em
(none)
50 mg/mL N/A N/A EtOH –20°C
Gentamycin Gm PURPLE 7.5 mg/mL 3 μg/mL (2500X) 15 μg/mL (500X) ddH2O 4°C
Hygromycin Hy
(none)
5 mg/mL 20 μg/mL (250X) 20 μg/mL (250X) ddH2O 4°C
Kanamycin Km YELLOW 7.5 mg/mL 30 μg/mL (250X) N/A ddH2O 4°C
Nalidixic acid Na
(none)
5 mg/mL 30 μg/mL (1000X) N/A ddH2O 4°C
Neomycin Nm GREEN 25 mg/mL N/A 100 μg/mL (250X) ddH2O 4°C
Rifampicin Rm
(none)
50 mg/mL N/A 100 μg/mL (500X) DMF –20°C
Spectinomycin Sp ORANGE 20 mg/mL 80 μg/mL (250X) 80 μg/mL (250X) ddH2O 4°C
Streptomycin Sm BLUE 50 mg/mL N/A 200 μg/mL (250X) ddH2O 4°C
Tetracycline Tc RED 2.5 mg/mL 10 μg/mL (250X) 10 μg/mL (250X) EtOH 4°C
Triclosan
(none)
N/A N/A EtOH 4°C

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. Exceptions: Gentamycin, Nalidixic Acid, and Rifampicin.

DNA

10 mM dNTPs

  • Thaw and mix (stir) 100 mM stock solutions of dGTP, dATP, dTTP, dCTP (NEB; in freezer)
  • Master Mix:
  • Dispense 100-μL aliquots into 1.7-mL tubes

Note: Store in the –20°C freezer in the box marked "Other Enzymes"

10X Taq buffer

  • Label tubes: 10X Taq buffer
  • Thaw and mix (stir) "Thermo pol buffer" (NEB; in freezer)
  • Aliquot 375 μL of "Thermo pol buffer" into tubes—do not dilute!

Note: Store in the –20°C freezer in the box marked "Other Enzymes"

10X Pfx buffer

  • Label tubes: 10X Pfx buffer
  • Thaw and mix (stir) "Pfx50 10X PCR Buffer" (NEB; in freezer)
  • Aliquot 375 μL of "Pfx50 10X PCR Buffer" into tubes—do not dilute!

Note: Store in the –20°C freezer in the box marked "Other Enzymes"

8X DNA loading

DNA ladder

Note: use 5 μL per lane; 3kb band = 60 ng

Ethidium Bromide (10 mL)

  • 10 mL ddH20
  • 10 mg ethidium bromide

Note: use 8 μL per 50 mL gel

Media Additives

1 M Calcium Chloride (75 mL)

  • 75 mL ddH2O
  • 11.027 g calcium chloride dihydrate (CaCl2·2H2O)

Autoclave

2 M Magnesium Chloride (75 mL)

  • 75 mL ddH2O
  • 30.4965 g magnesium chloride hexahydrate (MgCl2·6H2O)

Autoclave

1 M Sodium Citrate (40 mL)

  • 40 mL ddH2O
  • 11.76 g sodium citrate tribasic dihydrate (Na3C6H5O7·2H2O)
  • pH to 5.7 with HCl

Sterile filter

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

1 M Copper (II) sulfate (50 mL)

  • 50 mL ddH2O
  • 7.98 g CuSO4

Sterile filter

10% SDS (75 mL)

  • 75 mL ddH2O
  • 7.5 g Sodium dodecyl sulfate (SDS)

Do not sterilize

20% SDS (75 mL)

  • 75 mL ddH2O
  • 15 g Sodium dodecyl sulfate (SDS)

Do not sterilize

30% Acrylamide (75 mL)

  • 75 mL ddH2O
  • 22.5 g acrylamide

40% Acrylamide (75 mL)

  • 75 mL ddH2O
  • 30 g acrylamide

10% Ammonium persulfate (75 mL)

  • 75 mL ddH2O
  • 7.5 g ammonium persulfate (APS)

5 M NaCl (75 mL)

  • 75 mL ddH2O
  • 21.92 g NaCl

Autoclave

80% Glycerol (75 mL)

  • 15 mL ddH2O
  • 60 mL glycerol

Microwave for 30-second intervals on high until dissolved
Autoclave

1 M DTT (5 mL)

  • 5 mL ddH2O
  • 771 mg Dithiothreitol (DTT; in fridge)

Aliquot 500 μL into Eppendorf tubes Store in –20°C

Orcinol spray

  • 5 mL ddH2O
  • 25 mL methanol (in fume hood)
  • 10 mL sulfuric acid (H2SO4 (in fume hood—be careful!)
  • 200 mg orcinol
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