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Team:ESBS-Strasbourg/14 October 2008
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* 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: | * 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) | ** 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) | ** (3) Ura3 (FI) + Rep_cyc46(FV) | ||
** (4) Ura3 (FI) + Rep_cyc16(FV) | ** (4) Ura3 (FI) + Rep_cyc16(FV) | ||
Latest revision as of 09:22, 15 October 2008
Contents |
DryLab
Modeling
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.
