Difference between revisions of "Team:Northwestern/stock"

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     </ul>
 
     </ul>
 
       <h2>Solutions</h2>
 
       <h2>Solutions</h2>
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       <h3>10X TAE</h3>
+
       <h3>10X TAE (for 1L):</h3>
 
       <ul>
 
       <ul>
 
       <li>900 mL dH<sub>2</sub>O</li>
 
       <li>900 mL dH<sub>2</sub>O</li>
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       </ul>
 
       </ul>
 
        
 
        
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       <h3>1% agarose</h3>
+
       <h3>1% agarose:</h3>
 
       <ul>
 
       <ul>
 
       <li>1 g agarose for every 100 mL of 1X TAE</li>
 
       <li>1 g agarose for every 100 mL of 1X TAE</li>
 +
      <li>Microwave until completely clear, boiling</li>
 +
      <ul><li>About 3 min, <strong>loosen the cap</strong> to allow vapor to escape</li></ul>
 +
      <li>Pour immediately</li>
 
       </ul>
 
       </ul>
 
        
 
        
−
       <h3>Culture plates</h3>
+
       <h3>Culture plates:</h3>
 
       <ul><li>1.5% (w/v) Bacto agar to LB</li>
 
       <ul><li>1.5% (w/v) Bacto agar to LB</li>
 
       <li>Autoclave to sterilize</li>
 
       <li>Autoclave to sterilize</li>

Latest revision as of 20:35, 16 October 2016

Protocols

Stock Reagents

Media Recipes

LB Media (for 1L):

  • 10 g tryptone peptone
  • 5 g yeast extract
  • 5 g NaCl
  • 1 mL 1N NaOH or 200 uL 5M NaOH
  • Add H2O to 1L
  • Autoclave to sterilize
  • Store at RT until used

TB Media (for 1L):

  • 12 g tryptone peptone
  • 24 g yeast extract
  • 4 mL glycerol
  • Add H2O to 1L
  • Adjust pH to 7.0 with 5M NaOH
  • Autoclave to sterilize
  • Store at RT until used
  • *TB media is used for dense colonies

SOC (for 1L):

  • 20 g tryptone peptone
  • 5 g yeast extract
  • 0.5 g NaCl
  • 47 mg KCl
  • 0.952 g MgCl2
  • 1.203 g MgSO4
  • 3.6 g glucose
  • Autoclave to sterilize
  • Store at RT until used

Solutions

10X TAE (for 1L):

  • 900 mL dH2O
  • 48.4 g Tris base
  • 11.4 mL glacial acetic acid
  • 3.72 g EDTA
  • H2O to 1L
  • Dilute 1:10 to use 1X TAE

1% agarose:

  • 1 g agarose for every 100 mL of 1X TAE
  • Microwave until completely clear, boiling
    • About 3 min, loosen the cap to allow vapor to escape
  • Pour immediately

Culture plates:

  • 1.5% (w/v) Bacto agar to LB
  • Autoclave to sterilize
  • Allow to cool in 50°C water bath until the bottle can be comfortably held
  • Add 1uL 1000X antibiotic for every mL of agar made
  • Mix gently to avoid making bubbles
  • Pour immediately into Petri dishes
  • Partially cover with lids to set at RT
  • Cover in aluminum foil, store lid-down at 4°C