Jonesma Lab:Protocols:ligation: Difference between revisions
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(New page: #Thaw reagents and store on ice #Combine the following reagents 2 μL Ligation Buffer 5 μL Digested backbone 11 μL Digested insert (smaller piece) 2 μL T4 DNA ligase #Incubate at ...) |
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1. Thaw reagents and store on ice | |||
2. Combine the following reagents | |||
2 μL Ligation Buffer | 2 μL Ligation Buffer | ||
5 μL Digested backbone | 5 μL Digested backbone | ||
11 μL Digested insert (smaller piece) | 11 μL Digested insert (smaller piece) | ||
2 μL T4 DNA ligase | 2 μL T4 DNA ligase | ||
3. Incubate at 22°C for 1 hr, 16°C O/N, 65°C for 10min (Use a PCR machine for this) | |||
4. Transform bacteria with your ligation | |||
Revision as of 01:23, 23 October 2015
1. Thaw reagents and store on ice 2. Combine the following reagents
2 μL Ligation Buffer
5 μL Digested backbone
11 μL Digested insert (smaller piece)
2 μL T4 DNA ligase
3. Incubate at 22°C for 1 hr, 16°C O/N, 65°C for 10min (Use a PCR machine for this) 4. Transform bacteria with your ligation
