From 2011.igem.org
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| | {{Kyoto_Background}} | | {{Kyoto_Background}} |
| | {{Kyoto_WikiDesign}} | | {{Kyoto_WikiDesign}} |
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| - | ===Ligation===
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| - | #Make 2µL of Mixture (the vector and the insert at 1 : 5-10) and Control (only the vector).
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| - | #Add 5µL Ligation High, 1µL T4 Kinase, and 7µL MilliQ to create a solution.
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| - | #Incubate at 16℃ for 30 min. If the colonies of E.coli transformed with the Control
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| - |
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| - | ===Restrictive Digestion===
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| - | <ol>
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| - | <li>Use EcoRI, XbaI, SpeI, PstI, (NEB)</li>
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| - | <li>Mix the following.
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| - | {|
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| - | |-
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| - | | Sample
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| - | | 5µL
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| - | |-
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| - | | 10xBuffer
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| - | | 1µL
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| - | |-
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| - | | Restriction Enzyme
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| - | | 0.1µL
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| - | |-
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| - | | MilliQ
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| - | | -3.9µL
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| - | |}
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| - | </li>
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| - | <li>Let stand for 2h at 37℃</li>
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| - | </ol>
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Latest revision as of 03:32, 5 October 2011
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