Team:ESBS-Strasbourg/6 October 2008

< back

WetLab
Katja:  goals for tomorrow:
 * 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
 * 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!