IGEM:PennState/Labbook/NoahJohnson/2007-8-2

August 2, 2007

• Ordered UV cuvettes
• Began growing up 6 3ml cultures of crp* in LB
• Will TELT dna prep them tonight or tomorrow

M9 Minimal Media (250mL each):

For ~250mL of media and a final 0.2% sugar concentration:

• 200mL sterile water

• 50mL of 5X M9 salts
  1. 12.8g Na2HP04*7H2O
  2. 3g KH2PO4
  3. 0.5g NaCl
  4. 1g NH4Cl
  5. Dissolve in 200mL deionized water and autoclave to sterilize
• 3.75g BactoAgar

Autoclave to sterilize and allow to cool until you can hold it for 2 seconds on the palm of your hand. Then add the following and pour into plates:

• 25mL sugar solution
  1. Add 0.5g glucose/xylose to 25mL sterile water
  2. Filter-sterilize before adding
  3. Makes a 12.5% sugar solution and a 0.2% overall glucose/xylose media
• 500 uL 1M MgSO4
  1. Dissolve 24.65g MgSO4*7H20 in 100mL H20
  2. Autoclave to sterilize
• 25uL 1M CaCl2
  1. Dissolve 14.7 g CaCl2*2H2O in 100mL H2O
  2. Autoclave to sterilize

I will make 250mL of each of the following:

• 0.2% glucose
• 0.2% glucose + 0.2% xylose
• 0.2% xylose
• 0.2* glycerol (- control)

Protocol for plates

• DNA prep on crp* cultures from this morning using TELT method

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