Team:Goettingen/week18-3

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#3 Chemoreceptor Library - 18th Week

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V08_29


Inoculation of LB-media for qrtPCR of different promoter constructs
  • Experiment:
    5 ml of LB-media with 50 µg/mL chloramphenicol were inoculated with E. coli strain BL21 haboring the vector pSB1C3 with following combinations of promoters and Tar receptor constructs:

    - Tar receptor under the control of constitutive promoter J23100
    - Tar receptor under the control of constitutive promoter J23104
    - Tar receptor under the control of constitutive promoter J23105
    - Tar receptor under the control of constitutive promoter J23106
    - Tar receptor under the control of constitutive promoter J23109
    - Tar receptor under the control of constitutive promoter J23112
    - Tar receptor under the control of constitutive promoter J23113
    - Tar receptor under the control of constitutive promoter J23114

    For more details click here.

    Cultures were incubated overnight at 37 °C, shaking.


V08_30


First Test for Isolaton of RNA, Synthesis of cDNA and qrtPCR approach
  • Experiment:
    RNA was isolated using Nuceleus Spin RNA II (Machery and Nagel).
    cDNA synthesis was performed using Reverse Transcription Kit (Qiagen).
    First approach of qrtPCR with different delutions of cDNA in H20
    For a detailed protocol click here.
  • Observations & Results
    cDNA which was 1:10 diluted in H20 showed the best results for qrtPCR approach of our promoter constructs.


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