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Team:KIT-Kyoto/kazukokekokko
From 2012.igem.org
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Latest revision as of 12:49, 25 September 2012
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TNFAIP3
PARTS BP reaction by Invitrogen gateway system1. PCR using primers containing the attB sequence. 2. Purify PCR product. 3. Add the following components to a 1.5 mL microcentrifuge tube at room temperature and mix:
4. Vortex BP ClonaseII enzyme mix briefly. Add 1 - 2 μL to the components above and mix well by vortexing and spin them down. 5. Incubate reaction at 25˚C for more than 1 hour. 6. Add 1 μL of 2 μg/μL Proteinase K solution and vortex briefly. 7. Incubate at 37˚C for 10 minutes. LR reaction by Invitrogen gateway system1. Add the following components to a 1.5 mL microcentrifuge tube at room temperature and mix:
2. Vortex LR ClonaseII enzyme mix briefly. Add 1 – 2 μL, to the components above and mix well by vortexing and spin down. 3. Incubate reaction at 25˚C for 16 hours. 4. Add 1 μL of 2 μg/μL Proteinase K solution and vortex briefly. 5. Incubate at 37˚C for 10 minutes. |
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