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V09_03
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.
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V09_04_
V09_04_1 Isolaton of RNA, Synthesis of cDNA and qrtPCR
- Experiment:
RNA was isolated using Nuceleus Spin RNA II (Machery and Nagel).
cDNA synthesis was performed using Reverse Transcription Kit (Qiagen).
qrtPCR with promoter constructs was performed, for a detailed protocol click here.
- Observations & Results
Strong variations of CP-values, so we obtained no useful data for calculation of expression rates. Experiment needs to be repeated.
V09_04_2 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.
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V09_05
Isolaton of RNA, Synthesis of cDNA and qrtPCR
- Experiment:
RNA was isolated using Nuceleus Spin RNA II (Machery and Nagel).
cDNA synthesis was performed using Reverse Transcription Kit (Qiagen).
qrtPCR with promoter constructs was performed, for a detailed protocol click here.
- Observations & Results
Strong variations of CP-values, so we obtained no useful data for calculation of expression rates. Experiment needs to be repeated.
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