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User:University of Washington/10 July 2008
From 2008.igem.org
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·CV13(Yep13)<br\> | ·CV13(Yep13)<br\> | ||
·pDPT51 | ·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<br> | ||
| + | //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]] | Back to [[Team:University_of_Washington/Notebook#Notebook]] | ||
Revision as of 20:39, 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
