"
User:University of Washington/10 July 2008
From 2008.igem.org
(Difference between revisions)
(→LuxR from AraC and TetR) |
(→LuxR from AraC and TetR) |
||
| Line 29: | Line 29: | ||
//Note: Electroporator 200 Ohms, 1.5 kV, with Tsy Media<br> | //Note: Electroporator 200 Ohms, 1.5 kV, with Tsy Media<br> | ||
//Note: Failed to put cells on ice while bringing them over to do the electroporation. Hopefully, they will still grow. | //Note: Failed to put cells on ice while bringing them over to do the electroporation. Hopefully, they will still grow. | ||
| - | + | *Tube 1: 45 ul XL1 Blue + 15 ul DNA#1 | |
| - | + | *Tube 2: 45 ul XL1 Blue + 2 ul DNA#1 | |
| - | + | *Tube 3: 45 ul XL1 Blue + 15 ul DNA#2 | |
| - | + | *Tube 4: 45 ul XL1 Blue + 2 ul DNA#2 | |
| - | + | *Tube 5: 45 ul XL1 Blue | |
- Plated 200 ul each on LB agar + Amp | - Plated 200 ul each on LB agar + Amp | ||
| - | |||
---- | ---- | ||
Back to [[Team:University_of_Washington/Notebook#Notebook]] | Back to [[Team:University_of_Washington/Notebook#Notebook]] | ||
Revision as of 20:41, 10 July 2008
Contents |
RP4 Conjugation
Conjugation protocol #1
·DH5α
·DH5α + S17-1 w/ RP4
·DH5α + naked RP4 DNA
·DH5α + amp resistant RFP plasmid
Lambda Red
·Synthesis tetR insert for KorA and trbA replacement
non-RP4 Conjugation
Mini Prep & Glycerol Stock
·CV13(Yep13)
·pDPT51
LuxR from AraC and TetR
- GFP E0240 grew, got some single colonies, stored in a fridge.
- Finished up QuikChange Mutagenesis: Dpn1 digestion.
- Did PCR purification with 30 ul EB Buffer.
- Transformed mutated AraC plasmids into XL1 Blue cells.
//Note: Electroporator 200 Ohms, 1.5 kV, with Tsy Media
//Note: Failed to put cells on ice while bringing them over to do the electroporation. Hopefully, they will still grow.
- Tube 1: 45 ul XL1 Blue + 15 ul DNA#1
- Tube 2: 45 ul XL1 Blue + 2 ul DNA#1
- Tube 3: 45 ul XL1 Blue + 15 ul DNA#2
- Tube 4: 45 ul XL1 Blue + 2 ul DNA#2
- Tube 5: 45 ul XL1 Blue
- Plated 200 ul each on LB agar + Amp
Back to Team:University_of_Washington/Notebook#Notebook
