IGEM:UC-Merced/2009/Notebook/E. hydro Express

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  P1 Phage
  P1 Phage
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1. Overnight culture did not grow. Possible reasons are: (1) The JW1228-1 culture used was old and found in the 4°C refrigerator. It is possible that the culture was no longer living. To address this possibility we will redo the procedure using frozen stock.  
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1. Overnight culture did not grow. Possible reason is: The JW1228-1 culture taken from the 4°C refrigerator was old and was probably no longer living. To address this possibility we will redo the procedure using frozen JW1228-1 stock.  
-
2. Prepared new overnight culture with frozen JW1228-1 glycerol stock (procedure carried out as on 10/13/12)
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2. Prepared new overnight culture with frozen JW1228-1 glycerol stock (following the procedure carried out on 10/13/12)
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    a. LB
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    a. LB
     b. Kanamycin stock 25mg/ml to add to strain JW1228-1
     b. Kanamycin stock 25mg/ml to add to strain JW1228-1
         C1V1=C2V2
         C1V1=C2V2
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       25000 ug/ml (V1) = 250 ug
       25000 ug/ml (V1) = 250 ug
       V1 = 0.01 ml = 10 uL  --> kanamycin to add to strain JW1228-1  
       V1 = 0.01 ml = 10 uL  --> kanamycin to add to strain JW1228-1  
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       Incubate kanomycin in LB broth until 10 am 10/15/12
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       Incubate kanomycin in LB broth until 10:30 am 10/15/12
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__NOEDITSECTION__
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Revision as of 21:44, 14 November 2012

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Lab Notebook

Beginning Tuesday November 13, 2012

10/13/2012 

 Generation P1 Phage Stock

  1. Prepared P1 Stock from Yale University
   a. LB
   b. Kanamycin stock 25mg/ml to add to strain JW1228-1
       C1V1=C2V2
      25000 ug/ml (V1) = 50ug/ml (5ml)
      C1 = 25000 ug/ml  --> kanamycin concentration
      C2 = 50 ug/ml   
      V2 = 5 ml broth
      25000 ug/ml (V1) = 250 ug
      V1 = 0.01 ml = 10 uL   --> kanamycin to add to strain JW1228-1 
     Incubate kanomycin in LB broth until 11am 10/14/12


10/14/12 

P1 Phage

1. Overnight culture did not grow. Possible reason is: The JW1228-1 culture taken from the 4°C refrigerator was old and was probably no longer living. To address this possibility we will redo the procedure using frozen JW1228-1 stock. 2. Prepared new overnight culture with frozen JW1228-1 glycerol stock (following the procedure carried out on 10/13/12) 

   a. LB
   b. Kanamycin stock 25mg/ml to add to strain JW1228-1
       C1V1=C2V2
      25000 ug/ml (V1) = 50ug/ml (5ml)
      C1 = 25000 ug/ml  --> kanamycin concentration
      C2 = 50 ug/ml   
      V2 = 5 ml broth
      25000 ug/ml (V1) = 250 ug
      V1 = 0.01 ml = 10 uL   --> kanamycin to add to strain JW1228-1 
     Incubate kanomycin in LB broth until 10:30 am 10/15/12
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