Fish2bCBR, Fish2CBL Primer (cytb-Fish), amplify 80 pb, Thomsen et al. 2012

PCR #5, Pag 92, Sept 26 2019

PCR reaction

• ~40 ng/ul DNA
• 0.5 ul primer (10um)
• 5 ul Hoststar Qiagen
• 12 ul total reaction

Cycling conditions (Thomsen et al. 2012)

• 7 min to 95°C
• 50 cycles: 94°C 30 s, 50°C 30 s, 72°C 20 s,
• 5 min to 72°C
• 4°C forever

I used new samples (9/11/2019 extraction). Nonspecific amplification. I will increase annealing temperature at 53°C like PCR #3.

FISH2CB, PCR