User:Karmella Haynes/Notebook/BioBrick cloning/2011/07/18

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07/18/11

  • ✓ Cultures/minipreps: KAH219-224/ pSB31 (4 each)



Minipreps
> Check with E/P digests

Reagent Volume Expected:
*pSB31 bands
**last frag + 300
1-4. 219/pSB31 1-4 = 4500*, 150*, 444, 1735, 532**
5-8. 220/pSB31 1-4 = 4500*, 150*, 444, 1190**
9-12. 221/pSB31 1-4 = 4500*, 150*, 444, 1490**
13-16. 222/pSB31 1-4 = 4500*, 150*, 636, 1735, 532**
17-20. 223/pSB31 1-4 = 4500*, 150*, 636, 1190**
21-24. 224/pSB31 1-4 = 4500*, 150*, 636, 1490**
13. pSB31 = 4500, 450
File:KAH 071811 gel1.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.


--> Only KAH220/pSB31 worked. Pick more colonies for other constructs (cultures, o/n).