Team:UTK-Knoxville/media

From 2014.igem.org

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<h2>MEDIA</h2>
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<h3 id="top">M9 no Carbon</h3><hr>
<h3 id="top">M9 no Carbon</h3><hr>
<p>For 1L:</p>
<p>For 1L:</p>

Latest revision as of 21:16, 15 July 2014

MEDIA


M9 no Carbon


For 1L:

  • 100mL 10X M9 Salts
  • 1mL 1mg/mL thiamine HCl
  • 50μL Trace Metals Solution
  • 100μL 1M CaCl2 (add last, while stirring)
  • Filter Sterilize
  • Glucose is added as appropriate from stock either into culture tubes or into media
  • Add appropriate antibiotics to culture

10X M9 Salts


  • 67.8g/L Na2HPO4
  • 30g/L KH2PO4
  • 5g/L NaCl
  • 10g/L NH4Cl
  • Filter sterilize

M9 Plates


For 500mL:

  • Autoclave 15g/L agar with ~448mL water
  • Cool Down
  • Add 50mL 10X M9 Salts
  • 0.5mL 1M MgSO4
  • 0.5mL 1mg/mL thiamine HCl
  • 25μL Trace Metals Solution
  • 50μL 1M CaCl2
  • Add glucose from stock and appropriate antibiotics

M9 Medium+20% Sucrose


For 1L:

  • Add 200g sucrose while heating and mixing with hotplate
  • Turn off heat, continue stirring
  • 4g/L glucose (to aid growth)
  • 100mL 10X M9 Salts
  • 1mL 1M MgSO4
  • 0.5mL 1mg/mL thiamine HCl
  • 50μL Trace Metals Solution
  • 100μL 1M CaCl2
  • Filter Sterilize
  • Add appropriate antibiotics as necessary

LB


For 1L:

  • Add 25g of LB broth and mix well with 1 liter H2O
  • Autoclave to sterilize

UTK