Difference between revisions of "840:153g:Projects/project23/2012/10/26"

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(Site-directed Mutagenesis)
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==Site-directed Mutagenesis==
 
==Site-directed Mutagenesis==
* On Tuesday, we ran the gel for last week's PCR products.  This was done on a dual-lane chamber, loaded as follows:
+
*On Friday, we ran Thursday's PCR products on a 1.8% Agarose gel.  The samples were loaded as follows:
  
  
*Top Lane:
+
*Upper lane
#100 bp ladder
+
#100bp ladder
#P+55
+
#Non-primered set without template
#P+60
+
#Upstream fragment without template
#P+65
+
#Primered set without template
#Positive Control
+
#Downstream fragment without template
#P-55
 
#P-60
 
#P-65
 
 
#100 bp ladder
 
#100 bp ladder
 +
#Downstream fragment
 +
#Primered set
 +
#Upstream fragment
 +
#Non-primered set
  
  
*Bottom Lane:
+
*Lower lane
#100 bp ladder
+
#Positive control
#N+55
 
#N+60
 
#N+65
 
#Positive Control
 
#N-55
 
#N-60
 
#N-65
 
 
#100 bp ladder
 
#100 bp ladder
 +
*All other lanes are replications of primered set
  
  
*We ran these on a 1% agarose gel for 30min at 120V
+
*Results:
 +
*Upper lane, wells 1-5
  
  
*Results:
 
  
  
 +
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__NOTOC__
 
__NOTOC__
 
[[category:OWWLabNotebookV1]]
 
[[category:OWWLabNotebookV1]]

Revision as of 15:26, 29 October 2012

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Site-directed Mutagenesis

  • On Friday, we ran Thursday's PCR products on a 1.8% Agarose gel. The samples were loaded as follows:


  • Upper lane
  1. 100bp ladder
  2. Non-primered set without template
  3. Upstream fragment without template
  4. Primered set without template
  5. Downstream fragment without template
  6. 100 bp ladder
  7. Downstream fragment
  8. Primered set
  9. Upstream fragment
  10. Non-primered set


  • Lower lane
  1. Positive control
  2. 100 bp ladder
  • All other lanes are replications of primered set


  • Results:
  • Upper lane, wells 1-5