User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/10/28: Difference between revisions
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= | =Colony PCR of [http://openwetware.org/wiki/User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli/2013/10/25 Golden Gate Assembly] of BD003 and BD004= | ||
* Pick 10 colonies of each plate and grow in 2 mL LB AMP broth for 6 hours. | |||
* Spin down the cells for 3 minutes in microcentrifuge with top speed. | |||
* Resuspend the pallet in 100μL dH2O. | |||
* Incubate the cells in 98°C for 5 minutes. | |||
* Spin down the cells for 3 minutes in microcentrifuge with top speed. | |||
* Take the supernatent as DNA template for PCR reactions. | |||
* PCR colonies at: 95°C for 3 min. , [95°C 3 min, 45°C 45 sec., 72°C 3 min.] ×35, 72°C 6min. 4°C ∞. | |||
Revision as of 20:04, 28 October 2013
 PcTF Subcloning in E. coli
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Colony PCR of Golden Gate Assembly of BD003 and BD004
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