Team:ESBS-Strasbourg/14 October 2008

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WetLab
Sandra & Katja:
 * In the hope to get a construct, which is transformable in to yeast, meaning which has an auxotrophic marker, we made the following ligations and transformations:
 * t (1) Ura3 (FI) + Rep_cyc100(=LexAo8_cyc100_CFP_Linker_NLS_adh1term) (FV)
 * (2) Ura3 (FI) + Rep_cyc73(FV)
 * (3) Ura3 (FI) + Rep_cyc46(FV)
 * (4) Ura3 (FI) + Rep_cyc16(FV)
 * t (5) Ura3 (FI) + tag_NLS(FV)
 * (6) Ura3 (FI) + tag_hsl1(=Gal1_CFP_Linker_hsl1_adh1term)(FV)
 * t (7) Ura3 (FI) + pSB1A2(=p1010 digested with EcoRI and SpeI) (old)(FV)
 * t (8) Ura3 (FI) + pSB1A2 (new)(FV)
 * t(11) Rep_cyc100 (BV) + Leu2 (BI)
 * (12) Rep_cyc73 (BV) + Leu2 (BI)
 * (13) Rep_cyc46 (BV) + Leu2 (BI)
 * (14) Rep_cyc16 (BV) + Leu2 (BI)
 * (15) tag_NLS (BV) + Leu2 (BI)
 * (16) tag_hsl1 (BV) + Leu2 (BI)
 * t(17) pSB1A2 (=p1010 digested with XbaI and PstI) (BV - old) + Leu2 (BI)
 * t(18) pSB1A2 (BV - new) + Leu2 (BI)
 * (28) pSB1A2 (BV - new) + Leu2 (BI - old)
 * As negativ controlls we ligated and transformed:
 * (A) pSB1A2 (FV - old)
 * t (B) pSB1A2 (BV - old)
 * (C) pSB1A2 (FV - new)
 * (D) pSB1A2 (BV - new)
 * (E) water

The constructs marked t were transformed with heat shock into the competent TOP10 cells lefted. All other transformation have been done with a chemical transformation methode using chemical competent dhalpa cells.