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 20:46, 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 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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