User:Karmella Haynes/Notebook/BioBrick cloning/2015/05/06: Difference between revisions

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| dH<sub>2</sub>O || 8.5
| dH<sub>2</sub>O || 8.5
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| &nbsp; || 15 μL --> 37°C/ ~15 min.
| &nbsp; || 15 μL  
|}
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--> 37°C/ ~10 min.




Revision as of 18:41, 6 May 2015

Karmella's BioBrick Cloning <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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05/05/15

  • Ryan - minipreps & digests


Minipreps

  • Use Sigma GenElute; elute with 75 μL elution solution
  • promoter/Regulator/MRV colonies
  • Cut w/ KpnI/EcoRI - BbsI drop-in of promoter destroys KpnI site; empty vector will produce 2 bands, successful clones will be linearized (1 cut)
  1. pAub-AubR/MRV
  2. pBja-BjaR/MRV
  3. pBra-BraR/MRV
  4. pRpa-RpaR/MRV


Reagent Volume Expected:1,2. pAub/AubR/MRV = 4240
3,4. pBja/BjaR/MRV = 4099
5,6. pBra/BraR/MRV = ###
7,8. pRpa/RpaR/MRV = 4116
Empty vector = ~4000, 971 		Hover name
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 3.0 μL
10X buffer 1.5
enzyme 1 1.0
enzyme 2 1.0
dH2O 8.5
  15 μL

--> 37°C/ ~10 min.


  • Measure [DNA]
Sample OD 260 260/280 ng/μL
1. 0.465 1.913 465.4


Conclusions

  • tba



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