User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/10/28: Difference between revisions

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* Take the supernatent as DNA template for PCR reactions.  
* 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 ∞.
* 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 ∞.
[[Image:Image:20131029 103655 (3).jpg '''Colony PCR for BD004 all the ''' ]]

Revision as of 17:42, 29 October 2013

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Colony PCR of 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 ∞.

File:Image:20131029 103655 (3).jpg Colony PCR for BD004 all the

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