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User:University of Washington/26 June 2008
From 2008.igem.org
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- Yeast transforming media 50% PEG, 10x LiAc, and 10x TE were obtained. | - Yeast transforming media 50% PEG, 10x LiAc, and 10x TE were obtained. | ||
| - | - Isolated yeast colonies were inoculated in 5 mL of YEPD media and incubated overnight at 30 degrees Celsius in a rotator. | + | - Isolated BY4741 yeast colonies were inoculated in 5 mL of YEPD media and incubated overnight at 30 degrees Celsius in a rotator. |
== Conjugation Plasmid == | == Conjugation Plasmid == | ||
Revision as of 19:06, 27 June 2008
Contents |
Yeast Shuttle Plasmid
- Yeast growth media YEPD was obtained.
- Yeast transforming media 50% PEG, 10x LiAc, and 10x TE were obtained.
- Isolated BY4741 yeast colonies were inoculated in 5 mL of YEPD media and incubated overnight at 30 degrees Celsius in a rotator.
Conjugation Plasmid
Mini Preps
·C600+RP4 construct
Glycerol Stocks
·C600+RP4
·S17-1
LuxR from AraC and TetR
- Got 1 colony for TetR, 24 for AraC
- Grew culture for Glycerol Stock and Miniprep
LuxR from pLac
- Got 13 colonies for R0010, and 20 colonies for J04430. I0462 failed to grow.
- Grew cultures of R0010 and J04430 for glycerol stock. I0462 transformed e.coli sat out overnight in non-selective media but were plated again on Amp anyway to see if anything would grow.
- Four colonies from the J04430 plate were selected, and re-plated on Amp agar for later use.
BioBrick Promoter Measurements
- Spotted filter paper sections were removed for the GFP reporter gene contained in a plasmid (pSB1A2-E0240), the standard backbone plasmid containing the ccdb toxin (pSB3K3-P1010), the fully-assembled standard reference promoter-RBS-GFP gene-backbone plasmid (I20260), and three different preassembled promoter test constructs with (hopefully) variable transcriptional strengths (J23150, J23151, J23102).
- Filter paper sections were soaked in 5 uL of deionized water for one hour, then centrifuged at 14,000 RPM for 3 minutes.
- The centrifuged plasmids were then stored at 4 degrees Celsius until arrival of the TOP10 E. coli strain.
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