GEMSTONE Research Data: PCR Images and Protocols

Tarih: 11.03.24 Kit/Enzyme: Vazyme 2 x Phanta Flash PCR ID: 20627 Generic Cre PCR Mix Sample DNA Reagent μlSample Name Conc. Volume in PCR Mix dH2O y N1 4,44 ng/μl 4,5 μl 2 x Phanta Flash Master Mix 12,5 N2 3,54 ng/μl 5,65 μl 25mM Mg WT1 20 ng/μl 1 μl 5M Betaine WT2 20 ng/μl 1 μl DMSO 10mM dNTP Taq 10 μM TG Primer_F 1 10 μM TG Primer_R 1 10 μM IPC Primer_F 1 10 μM IPC Primer_R 1 Alq 16,5 + y x ng/μl DNA 20 Primer Samples PCR Condition Program: TG forward: GCG GTC TGG CAG TAA AAA CTA TC N1 initial denaturation: 98°C 30 sec TG reverse: GTG AAA CAG CAT TGC TGT CAC TT N2 35 cycle IPC forward: CAC GTG GGC TCC AGC ATT WT1 denaturation: 98°C 30 sec IPC reverse: TCA CCA GTC ATT TCT GCC TTT G WT2 annealing: 60 °C 5 sec extension: 72°C 1 sec Product Size: IPC = 74 bp Final Extension 72°C 1 min Tg = 102 bp Notes 3% agarose gel / 110 V - 60 mins Tarih: 13.03.24 Kit/Enzyme: Vazyme 2 x Phanta Flash PCR ID: 27807 Snca flox PCR Mix Sample DNA Reagent μlSample Name Conc. Vol in Mix dH2O y S1 5,3 ng/μl 3,77 μl 2 x Phanta Flash Master Mix 12,5 S2 3,92 ng/μl 5,1 μl 25mM Mg WT1 20 ng/μl 1 μl 5M Betaine WT2 20 ng/μl 1 μl DMSO 10mM dNTP Taq 12,5 μM Primer F 1 12,5 μM Primer R 1 Alq 16,5 + y x ng/μl DNA 20 Primer Samples PCR Condition Program: Forward: AAG AGC TAG TGG TGG GCA GA S1 initial denaturation: 98°C 30 sec Reverse: GCT GGG CAC AGT GTT GAT TG S2 35 cycle WT1 denaturation: 98°C 30 sec WT2 annealing: 60 °C 5 sec extension: 72°C 1 sec Product Size: Mutant = ~440 bp Final Extension Heterozygote = ~440 bp & 390 bp 72°C 1 min Wild type = 390 bp Notes 2% agarose gel / 110 V - 85 mins