User:Karmella Haynes/Notebook/BioBrick cloning/2011/05/30

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05/30/11

  • ✓ Miniprep digests: check miR sensor set (re-transformed)



Minipreps
> Check with E/P digests

Reagent Volume Expected:
MV2 = 4529
1,2. FC1 = 8063
3,4. FC2 = 8255
5,6. FC3 = 7991
7. FC4 = 8183
8. FC31 = 3052
9. FC32 = 2980
10. FC33 = 3244
11. FC34 = 3172
12,13. KAH118 = 7930
14,15. KAH119 = 8002
16. KAH100 = 3085
17. KAH101 = 3157
18,19. KAH121 = 8102
20,21. KAH122 = 8174
22. KAH124 = 8174
23,24. KAH125 = 8246
File:KAH 053011 gel1.tif
15 μL/lane; 1% agarose


File:KAH 053011 gel2.tif
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.

-> Note: all worked except for FC1, KAH118, KAH119, KAH124. Use retransformations (#1, 2, 12, 13, 14, 15, 22) to grow 5mL cultures and try again.
--> Result: Miniprep digests do not look good for FC1, KAH118, KAH119, but spec readings show ~160 ng/μL. Try one of each for transfection to see what happens.
-> Spec readings show good concentrations for all others. Keep for U2OS transfectons.
-> Made streak plate from 2 mL cultures yesterday.