Difference between revisions of "IGEM:UC-Merced/2009/Notebook/E. hydro Express"

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Line 29: Line 29:
 
10/13/2012
 
10/13/2012
  
   Phage
+
   Generation P1 Phage Stock
  
   1. Prepared Stocks
+
   1. Prepared P1 Stock from Yale University
 
     a. LB
 
     a. LB
     b. Kanomycin to add to strain JW1228-1
+
     b. Kanamycin stock 25mg/ml to add to strain JW1228-1
 
         C1V1=C2V2
 
         C1V1=C2V2
       25000 ug (V1) = 50ug (5ml)
+
       25000 ug/ml (V1) = 50ug/ml (5ml)
       C1 = 25000 ug  --> kanomycin concentration
+
       C1 = 25000 ug/ml --> kanamycin concentration
       C2 = 50 ug,   V2 = 5 ml broth
+
       C2 = 50 ug/ml    
       25000 ug (V1) - 250 ug (ml)
+
      V2 = 5 ml broth
       V1 = 0.01 ml = 10 uL  --> kanomycin to add to strain JW1228-1  
+
       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
 
       Incubate kanomycin in LB broth until 11am 10/14/12
  
Line 45: Line 46:
 
10/14/12
 
10/14/12
  
  Phage
+
P1 Phage
  
1. Overnight culture did not grow.  
+
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.  
2. Prepared new stock with frozen bacteria (procedure carried out as on 10/13/12)
+
2. Prepared new overnight culture with frozen JW1228-1 glycerol stock (procedure carried out as on 10/13/12)
    a. LB
+
    a. LB
     b. Kanomycin to add to strain JW1228-1
+
     b. Kanamycin stock 25mg/ml to add to strain JW1228-1
 
         C1V1=C2V2
 
         C1V1=C2V2
       25000 ug (V1) = 50ug (5ml)
+
       25000 ug/ml (V1) = 50ug/ml (5ml)
       C1 = 25000 ug  --> kanomycin concentration
+
       C1 = 25000 ug/ml --> kanamycin concentration
       C2 = 50 ug,   V2 = 5 ml broth
+
       C2 = 50 ug/ml    
       25000 ug (V1) - 250 ug (ml)
+
      V2 = 5 ml broth
       V1 = 0.01 ml = 10 uL  --> kanomycin to add to strain JW1228-1  
+
       25000 ug/ml (V1) = 250 ug
       Incubate kanomycin in LB broth until 11am 10/14/12
+
       V1 = 0.01 ml = 10 uL  --> kanamycin to add to strain JW1228-1  
 +
       Incubate kanomycin in LB broth until 10 am 10/15/12
  
 
      
 
      

Revision as of 17:46, 14 November 2012

<!-- sibboleth --><div id="lncal1" style="border:0px;"><div style="display:none;" id="id">lncal1</div><div style="display:none;" id="dtext"></div><div style="display:none;" id="page">IGEM:UC-Merced/2009/Notebook/E. hydro Express</div><div style="display:none;" id="fmt">yyyy/MM/dd</div><div style="display:none;" id="css">OWWNB</div><div style="display:none;" id="month"></div><div style="display:none;" id="year"></div><div style="display:none;" id="readonly">Y</div></div>

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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 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. 2. Prepared new overnight culture with frozen JW1228-1 glycerol stock (procedure carried out as 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 am 10/15/12




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