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

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(Autocreate 2013/10/28 Entry for User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli)
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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=
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* Pick 10 colonies of each plate and grow in 2 mL LB AMP broth for 6 hours.
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* Spin down the cells for 3 minutes in microcentrifuge with top speed.
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* Resuspend the pallet in 100μL dH2O.
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* Incubate the cells in 98°C for 5 minutes.
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* Spin down the cells for 3 minutes in microcentrifuge with top speed.
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* Take the supernatent as DNA template for PCR reactions.
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* 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 22:04, 28 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 ∞.
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