"
Team:Brown/Notebook/July18
From 2010.igem.org
Sunday, July 18 2010
Results of 7/17 test of competent cells:
XL1B
- With 100 pg of RFP plasmid: 16 colonies
- With 1 ng of RFP plasmid: ~150 colonies
- With 10 ng of RFP plasmid: >800 colonies
XL1B
- With 100 pg of empty pNoTat: ~50 colonies
- With 1 ng of empty pNoTat: ~180 colonies
- With 10 ng of empty pNoTat: a lot of colonies (thousands).
Preparation of Tat BioBricks from registry:
pAraC = R0080
- Plate 1: 12E
AraC = C0080
- Plate 1: 14L
- Kan resistance
LacI = I732100
- Plate 2: 10E
Terter = B0015
- Plate 1: 23L
Punched hole, added 10 µl of ddH2O.
Used 3 mL of rehydrated DNA to transform XL1.
Plated and incubate at 37°C overnight.
7:00 PM: Removed 8 aliquots of competent cells (Xl1B) from yesterday and let them thaw on ice.
7:04 PM: Removed 3 null plates, 3 amp plates, 3 kan plates, and 3 tet plates from the fridge.
7:22 PM: Diluted BBa_K19005 (150 ng/µl) to 149 µl dH2O, results in 1 ng/µl concentration DNA.
Also, diluted Sosnick vector TrpR by diluting 1 µl of plasmid at 393 ng/µl into 392 µl H2O for a final concentration of 1 ng/µl.
Tat Parts List
- pAraC__Bba_R0080__’10 Plate 1__12E__pSB1A2
- RBS-ChFP-Terter__Bba_J06702__’10 Plate 2__8E__pSB1A2
- AraC__Bba_C0080__’10 Plate 1__14L__pSB2K3
- LacI__Bba_I732100__’10 Plate 2__10E__pSB1A3
- Terter__Bba_B0015__’10 Plate 1__23L__pSB1AK3
- pLac-RBS-CFP-Terter__Bba_J04421__’08 Plate 1__4G__pSB1A2
Accidentally diluted stock of BBa_K19005 with 149 µl dH2O, re-nanodropped and got a reading of 18.4 ng/µl. Added 1 µl of this to 17.4µl dH2O to make 1 ng/µl stock.
8:07 PM: Added 10 ng DNA to each aliquot and incubated for 20 minutes.
Later: Finished cell transformation protocol, placed in incubator.
