User:Karmella Haynes/Notebook/BioBrick cloning/2015/05/29

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mm/dd/yy

  • David Tze - minpreps & digests
  • Ben - sequencing order
  • Cas-tone - sequencing order


David Tze - minpreps & digests

  • Minipreps
    • Sigma GenElute kit, elute w/ 75 μL elution sln.
  • Digests
    • Check with E/S digests; cannot use PstI because PLrigid & PLrigid4 contain PstI sites
Reagent Volume Expected:
1. DT001 = size
2. DT002 = size
3. DT003 = size
4. DT004 = size
Empty vectors = ~2600, ~400 - 600 		Hover name
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 2.0 μL
10X buffer 1.5
EcoRI 1.0
SpeI 1.0
dH2O 9.5
  15 μL --> 37°C/ ~15 min.

Sequencing orders

  1. DBN007_1 - VF2
  2. DBN007_1 - VR
  3. DBN007_2 - VF2
  4. DBN007_2 - VR
  5. H3140_1 - CMV_f
  6. H3140_1 - DD123_r
  7. H3140_2 - CMV_f
  8. H3140_2 - DD123_r
  9. H3180_1 - CMV_f
  10. H3180_1 - DD123_r
  11. H3180_2 - CMV_f
  12. H3180_2 - DD123_r





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