Team:Cambridge/Gibson/Protocol

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(Difference between revisions)
(Step 3: PCR)
(Step 3: PCR)
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{{:Team:Cambridge/Templates/RightImage|image=Phusion.jpg|caption=Phusion Polymerase}}
{{:Team:Cambridge/Templates/RightImage|image=Phusion.jpg|caption=Phusion Polymerase}}
PCR is a bit of a dark art, but we have found that these general principles have served us well over the summer.
PCR is a bit of a dark art, but we have found that these general principles have served us well over the summer.
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{|class="wikitable"
 
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|-
 
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|Step
 
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|Temp
 
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|Time
 
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|-
 
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|1:Initial Melting
 
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|98°C
 
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|30s
 
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|-
 
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|2:Melting
 
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|98°C
 
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|10s
 
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|-
 
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|3:Annealing
 
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|T<sub>m</sub>°C
 
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|15s
 
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|-
 
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|4:Elongation
 
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|72
 
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|45s per kb DNA
 
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|-
 
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|5:GoTo step 2
 
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|
 
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|30 times
 
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|-
 
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|6:Final Elongation
 
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|72°C
 
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|7m30
 
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|-
 
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|7:Final Hold
 
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|4°C
 
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|∞
 
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|}
 
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<html>
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Revision as of 19:36, 24 October 2010