User:Karmella Haynes/Notebook/BioBrick cloning/2010/04/15

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04/15/10

  • ✓ Minipreps: KAH142, 146, 147/V0201 (20 total)
  • ✓ Cultures: 6-hour 2 mL cultures for KAH154-159/MV2 (2 each)
  • ✓ Minipreps: KAH154-159/MV2 (12 total)
  • ✓ Cultures: KAH142(10), 146/V0201(10)
  • ✓ Cultures: V0201 and MV2 (2 each)



Minipreps
> Check with XbaI digests

Reagent Volume Expected:
V0201 band = 4516
Insert band = BB size + 641 bp
1-6. KAH142/V0201 = 5212
7-12. KAH146/V0201 = 7027
13-20. KAH147/V0201 = 8773
XbaI digests 3/15/10
15 μL/lane; 1% agarose
DNA(plasmid) 2.0 μL
10X buffer 1.5
XbaI 1.0
dH2O 10.5
  15 μL

--> 37°C/ ~15 min.

--> Success: KAH147/V0201
--> Pick more colonies for KAH142, 146/V0201


Minipreps 2
> KAH154-159/MV2
> Check with E/P digests

Reagent Volume Expected:
MV2 band = 4529
1,2. KAH154/MV2 = 1724
3,4. KAH155/MV2 = 1724
5,6. KAH156/MV2 = 1724
7,8. KAH157/MV2 = 1724
9,10. KAH158/MV2 = 2688
11,12. KAH159/MV2 = 2691
E/P digests 4/15/10
15 μL/lane; 1% agarose
DNA(plasmid) 2.0 μL
10X buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 9.5
  15 μL

--> 37°C/ ~15 min.


--> Pick 4 more colonies for KAH157/MV2



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