Noi:LabNotes/2010-10-21: Difference between revisions

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(Created page with '==Probe prep for the DMR220k_S4 and S5== (followed Dr. Zhang's conditions) ===Production PCR=== 10nM 1st …')
 
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===Production PCR===
===Production PCR===
                                                                        
                                                                        
10nM 1st round amplicon      0.2ul                 
    10nM 1st round amplicon      0.2ul                 
     2x Kapa SYBG qPCR Master Mix  50ul         
     2x Kapa SYBG qPCR Master Mix  50ul         
     100uM pAP1V61U              0.4ul                   
     100uM pAP1V61U              0.4ul                   
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     95C 30sec -> (95C 3sec -> 55C 30sec-> 60C 20sec) x 16 -> 60C 2min -> 15C hold.
     95C 30sec -> (95C 3sec -> 55C 30sec-> 60C 20sec) x 16 -> 60C 2min -> 15C hold.
     Ethanol precipitation; re-purified with twelve Qiaquick columns, each eluted with 40ul EB.
     Ethanol precipitation; re-purified with twelve Qiaquick columns, each eluted with 40ul EB (total vol: 12 x 40 = 480ul).
      
      


                                                                         DMR220k_S4                                            DMR220k_S5
                                                                         DMR220k_S4                                            DMR220k_S5
     DNA concentration (ng/ul)                                                                    
     DNA concentration (ng/ul)                                                    
     O.D.260/280
     O.D.260/280                                    
     Yield: concxtotal vol.
     Yield: concxtotal vol.

Revision as of 14:51, 22 October 2010

Probe prep for the DMR220k_S4 and S5

(followed Dr. Zhang's conditions)

Production PCR

   10nM 1st round amplicon      0.2ul                
   2x Kapa SYBG qPCR Master Mix  50ul        
   100uM pAP1V61U               0.4ul                  
   100uM AP2V6                  0.4ul                      
   H2O                           49ul                          
   ===================================
                                100ul x 96
   95C 30sec -> (95C 3sec -> 55C 30sec-> 60C 20sec) x 16 -> 60C 2min -> 15C hold.
   Ethanol precipitation; re-purified with twelve Qiaquick columns, each eluted with 40ul EB (total vol: 12 x 40 = 480ul).
   
                                                                       DMR220k_S4                                             DMR220k_S5
   DNA concentration (ng/ul)                                                     
   O.D.260/280                                     
   Yield: concxtotal vol.