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Team:ESBS-Strasbourg/14 October 2008
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'''Sandra & Katja:''' | '''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: | * 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: | ||
| + | <pre> | ||
** 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) | ||
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** (D) pSB1A2 (BV - new) | ** (D) pSB1A2 (BV - new) | ||
** (E) water | ** (E) water | ||
| - | + | <pre> | |
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. | 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. | ||
Revision as of 09:19, 15 October 2008
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 <pre> 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. == General == == Quote of the day ==
