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

PCR #6, Pag 97, Oct 2 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, 53°C 30 s, 72°C 20 s,
• 5 min to 72°C
• 4°C forever

Fragment sizes were verified on 1% agarose gels stained with GelRed 4ul PCR product 4ul 1kb 100 v, 2 hrs.

Nonspecific amplification again. We will test new samples extracted with MACHEREY-NAGEL kit to obtain best results in DNA yield.

FISH2CB, PCR