User:Karmella Haynes/Notebook/BioBrick cloning/2010/04/06

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04/06/10

  • ✓ Assembly: KAH142 (re-build) (finished on 4/07/10)



Assembly

  1. KAH142: (1) KAH014(DPRE)/(E/S)/1740 + (2) KAH96/(E/X)/~7771

> Digests (Fermentas FD)

Reagent Volume   Hover name
30 μL/lane, 1% agarose
DNA (plasmid) up to 25 μL
10x buffer 3.0
enzyme 1 1.0
enzyme 2 1.0
dH2O ---
  30 μL --> 37°C/ ~30 min.


> Measure conc.'s

Sample OD260 260/280 ng/μL
1. KAH014 (E/S) 0.675 1.77 33.7
2. KAH96 (E/X) 1.476 1.83 73.8


> Ligations

Ligation Plate results (lig : neg crtl) mm/dd/yy
1. KAH014(E/S)/1740, 18 ng + KAH96(E/X)/7771, 40 ng KAH142 rebuilt >10:1 (Pick 4)
2. KAH96(E/X)/ 40 ng 2 colonies
  1 2
Insert DNA 0.5 ---
Vector DNA 0.5 0.5
2x lgn buf (Roche) 5.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 3.0 3.5
  10 μL 10 μL



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