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Team:Amsterdam/Notebook/Protocols/Ligations
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(Created page with "{{:Team:Amsterdam/Header}} =Ligations== After following our restriction digest protocol (which uses 500ng of DNA) you may follow these steps for ligation. 1. Add 11ul of dH20 2...")
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(Created page with "{{:Team:Amsterdam/Header}} =Ligations== After following our restriction digest protocol (which uses 500ng of DNA) you may follow these steps for ligation. 1. Add 11ul of dH20 2...")
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Revision as of 11:34, 27 July 2011
Ligations=
After following our restriction digest protocol (which uses 500ng of DNA) you may follow these steps for ligation.
1. Add 11ul of dH20
2. Add 2ul from each sample you will be ligating (destination plasmid, and part)
3. Add 2ul of T4 DNA Ligase Reaction Buffer
4. Add 1ul of T4 DNA Ligase
5. Mix well, and spin down.
6. Incubate for 30min at 16C and 20min at 80C to heat kill.
7. Use 2ul of ligation to transform into competent cells.
