Team:LMU-Munich/Notebook/Protocols/24 Colony PCR

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(Difference between revisions)
(New page: {{:Team:LMU-Munich/Templates/Page Header}} Colony PCR {| |- |Component |Volume |Final concentration |- |PCR Master Mix, 2X |50 µl |1x |- |upstream primer | | |- |downstream primer ...)
 
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|Component
|Component
|Volume
|Volume
-
|Final concentration
 
|-
|-
-
|PCR Master Mix, 2X
+
|PCR Master Mix
-
|50 µl
+
|2.5µl
-
|1x
+
-
|-
+
|-  
-
|upstream primer  
+
|primer forward (BioBrick)
-
|
+
|0.375µl
-
|
+
|-
|-
-
|downstream primer
+
|primer revers (BioBrick)
-
|
+
|0.375µl
-
|
+
-
 
+
-
|-
+
-
|DNA Template
+
-
|
+
-
|
+
|-
|-
|nuclease free water to final volume of
|nuclease free water to final volume of
-
|
+
|5µl
-
|
+
|}
|}
 +
 +
 +
Guidelines for Amplification by PCR
 +
 +
:A. Denaturation:
 +
::initial: 94°C 5'
 +
::subsequent: 30''
 +
:B. Annealing:
 +
::53°C 30''
 +
:C. Extension
 +
::72°C 1'30''
 +
::final: 8'
 +
:D. Refrigeration
 +
::16°C
 +
:E.Cycle Number
 +
::30 cycles
 +
 +
 +
{{:Team:LMU-Munich/Templates/Page Footer}}

Latest revision as of 12:47, 27 September 2010


Colony PCR

Component Volume
PCR Master Mix 2.5µl
primer forward (BioBrick) 0.375µl
primer revers (BioBrick) 0.375µl
nuclease free water to final volume of 5µl


Guidelines for Amplification by PCR

A. Denaturation:
initial: 94°C 5'
subsequent: 30
B. Annealing:
53°C 30
C. Extension
72°C 1'30
final: 8'
D. Refrigeration
16°C
E.Cycle Number
30 cycles


 

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