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Team:ESBS-Strasbourg/6 October 2008
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** Ura3 (FI with Pst1 site) + pSB1A2 (FV) | ** Ura3 (FI with Pst1 site) + pSB1A2 (FV) | ||
** Leu2 (BI with EcoRI site) + pSB1A2 (BV) | ** Leu2 (BI with EcoRI site) + pSB1A2 (BV) | ||
| + | ** Ura3 (FI with Pst1 site) + LexAo8_cyc100_Kozak_CFP_NLS_adhterm (''alias Reporter cyc100'') (FV) | ||
| + | ** Ura3 (FI with Pst1 site) + LexAo8_cyc16_Kozak_CFP_NLS_adhterm (''alias Reporter cyc16'') (FV) | ||
| + | ** Ura3 (FI with Pst1 site) + LexAo8_cyc43_Kozak_CFP_NLS_adhterm (''alias Reporter cyc43'') (FV) | ||
** Gal1 (BV)+ CFP_NLS_adhterm (BI) | ** Gal1 (BV)+ CFP_NLS_adhterm (BI) | ||
** Gal1 (BV)+ CFP_cln2_adhterm (BI) | ** Gal1 (BV)+ CFP_cln2_adhterm (BI) | ||
| Line 18: | Line 21: | ||
* Preparation of LB-media (2x150ml, 2x100ml) | * Preparation of LB-media (2x150ml, 2x100ml) | ||
* Start of ON of lexA_L, mCherry_L_VP16_L, cin8_adhterm | * Start of ON of lexA_L, mCherry_L_VP16_L, cin8_adhterm | ||
| - | + | <br\n> | |
''goals for tomorrow:'' | ''goals for tomorrow:'' | ||
* Verification of: | * Verification of: | ||
| - | ** Ura3 | + | ** Ura3 with Verf primers |
| - | ** Leu2 | + | ** Leu2 with Verf primers |
| - | ** Gal1_CFP_NLS_adhterm | + | ** Gal1_CFP_NLS_adhterm with XFP and Verf primers |
| - | ** Gal1_CFP_cln2_adhterm | + | ** Gal1_CFP_cln2_adhterm with XFP and Verf primers |
| - | ** Gal1_CFP_hsl1_adhterm | + | ** Gal1_CFP_hsl1_adhterm with XFP and Verf primers |
| - | ** Gal1_mCherry_L_lexA_L_VP16_L | + | ** Gal1_mCherry_L_lexA_L_VP16_L with XFP and adhterm primers |
| - | ** Gal1_lexA_L_VP16_L_mCherry_L | + | ** Gal1_lexA_L_VP16_L_mCherry_L with XFP and adhterm primers |
| + | ** Ura3_Rep cyc100 Verf For/ minP Rev (attended fragment: 120+~800+296+110=1330) | ||
| + | ** Ura3_Rep cyc16 Verf For/ minP Rev (attended fragment: 120+~800+296+110=1330) | ||
| + | ** Ura3_Rep cyc43 Verf For/ minP Rev (attended fragment: 120+~800+296+110=1330) | ||
| + | * Start of ON of all positive clones (+ save plates) | ||
| + | * Ligation of: | ||
| + | ** CFP (BV-ready) + cin8_adhterm (BI-have to be prepared) | ||
| + | ** lexA_L (BV-have to be prepared) + mCherry_L_VP16_L (BI-have to be prepared) | ||
== General == | == General == | ||
| + | |||
| + | There are only 10 aliquotes of competent TOP10 cells left! | ||
== Quote of the day == | == Quote of the day == | ||
Latest revision as of 19:00, 6 October 2008
Contents |
DryLab
Modeling
WetLab
Katja:
- Verification PCR for Leu2 and Ura3 parts -> negative
- Ligation and Transformation of:
- Ura3 (FI with Pst1 site) + pSB1A2 (FV)
- Leu2 (BI with EcoRI site) + pSB1A2 (BV)
- Ura3 (FI with Pst1 site) + LexAo8_cyc100_Kozak_CFP_NLS_adhterm (alias Reporter cyc100) (FV)
- Ura3 (FI with Pst1 site) + LexAo8_cyc16_Kozak_CFP_NLS_adhterm (alias Reporter cyc16) (FV)
- Ura3 (FI with Pst1 site) + LexAo8_cyc43_Kozak_CFP_NLS_adhterm (alias Reporter cyc43) (FV)
- Gal1 (BV)+ CFP_NLS_adhterm (BI)
- Gal1 (BV)+ CFP_cln2_adhterm (BI)
- Gal1 (BV)+ CFP_hsl1_adhterm (BI)
- Gal1 (BV)+ mCherry_L_lexA_L_VP16_L (BI) (Ligation by Sandra)
- Gal1 (BV)+ lexA_L_VP16_L_mCherry_L (BI) (Ligation by Sandra)
- Preparation of LB-media (2x150ml, 2x100ml)
- Start of ON of lexA_L, mCherry_L_VP16_L, cin8_adhterm
goals for tomorrow:
- Verification of:
- Ura3 with Verf primers
- Leu2 with Verf primers
- Gal1_CFP_NLS_adhterm with XFP and Verf primers
- Gal1_CFP_cln2_adhterm with XFP and Verf primers
- Gal1_CFP_hsl1_adhterm with XFP and Verf primers
- Gal1_mCherry_L_lexA_L_VP16_L with XFP and adhterm primers
- Gal1_lexA_L_VP16_L_mCherry_L with XFP and adhterm primers
- Ura3_Rep cyc100 Verf For/ minP Rev (attended fragment: 120+~800+296+110=1330)
- Ura3_Rep cyc16 Verf For/ minP Rev (attended fragment: 120+~800+296+110=1330)
- Ura3_Rep cyc43 Verf For/ minP Rev (attended fragment: 120+~800+296+110=1330)
- Start of ON of all positive clones (+ save plates)
- Ligation of:
- CFP (BV-ready) + cin8_adhterm (BI-have to be prepared)
- lexA_L (BV-have to be prepared) + mCherry_L_VP16_L (BI-have to be prepared)
General
There are only 10 aliquotes of competent TOP10 cells left!
