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A Anhang A.1 PCR-Details Im folgenden Abschnitt s<strong>in</strong>d für die im Abschnitt 4.1.1 beschriebenen PCRs die Reaktionsansätze sowie die entsprechenden PCR-Progamme im Detail aufgeführt. A.1.1 PCRs <strong>zu</strong>r 3’-UTR Amplifikation Tabelle A.1: PCR-Details <strong>zu</strong>r Amplifikation der AGTR1 3’-UTR PCR-Ansatz f.c. PCR-Programm cDNA Bibliothek 0,2 µg 1. 1 × Denaturierung 98 °C 180 s 5×Phusion HF Puffer 1 × 2. 35 × Denaturierung 98 °C 10 s dNTPs 0,8 mM Anneal<strong>in</strong>g 64 °C 20 s AGTR1 FW SpeI 2 0,5 µM Extension 72 °C 30 s AGTR1 RV SacI 2 0,5 µM 3. 1 × f<strong>in</strong>ale Extension 72 °C 450 s Phusion Pol 1 u f.v. 50 µl Tabelle A.2: PCR-Details <strong>zu</strong>r Amplifikation der ESR1 3’-UTR PCR-Ansatz f.c. PCR-Programm cDNA aus MCF-7 RNA 0,125 µg 1. 1 × Denaturierung 98 °C 180 s 5×Phusion HF Puffer 1× 2. 35 × Denaturierung 98 °C 30 s dNTPs 0,8 mM Anneal<strong>in</strong>g 70 °C 30 s ESR1 FW SpeI 0,5 µM Extension 72 °C 60 s ESR1 RV MluI 0,5 µM 3. 1 × f<strong>in</strong>ale Extension 72 °C 450 s Velocity bzw. Precisor Pol 1 u f.v. 50 µl 165