IGEM:PennState/Labbook/NoahJohnson/2007-8-2: Difference between revisions

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<h2>M9 Minimal Media (250mL each):</h2>
<h2>M9 Minimal Media (250mL each):</h2>
**0.2% glucose
**0.2% glucose + 0.2% xylose
**0.2% xylose
**0.2* glycerol (- control)


For 250mL of media:
For ~250mL of media and a final 0.2% sugar concentration:
*200mL sterile water  
*200mL sterile water  
*25mL sugar solution
 
*#Add 0.5g glucose/xylose to 25mL sterile water
*#Filter-sterilize before adding
*#Makes a 12.5% sugar solution and a 0.2% overall glucose/xylose media
*50mL of 5X M9 salts
*50mL of 5X M9 salts
*#12.8g Na2HP04*7H2O
*#12.8g Na2HP04*7H2O
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*#1g NH4Cl
*#1g NH4Cl
*#Dissolve in 200mL deionized water and autoclave to sterilize
*#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
*#Add 0.5g glucose/xylose to 25mL sterile water
*#Filter-sterilize before adding
*#Makes a 12.5% sugar solution and a 0.2% overall glucose/xylose media
*500 uL 1M MgSO4
*500 uL 1M MgSO4
*#Dissolve 24.65g MgSO4*7H20 in 100mL H20
*#Dissolve 24.65g MgSO4*7H20 in 100mL H20
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*#Dissolve 14.7 g CaCl2*2H2O in 100mL H2O
*#Dissolve 14.7 g CaCl2*2H2O in 100mL H2O
*#Autoclave to sterilize
*#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)




[http://www.thelabrat.com/protocols/m9minimal.shtml Another protocol]
[http://www.madsci.org/~lynn/micro/techniques/pour_plates/MINagar.html Protocol for plates]
[http://www.madsci.org/~lynn/micro/techniques/pour_plates/MINagar.html Protocol for plates]


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


10:30-
10-

Latest revision as of 16:44, 2 August 2007

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

10-

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