Team:ESBS-Strasbourg/6 October 2008

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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!

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