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

PCR #1, Pag 60, Jul 23 2019

PCR reaction

• ~40 ng/ul DNA
• 1 ul primer (10um)
• Hoststar Qiagen 10 ul
• BSA 1 ul
• 25 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

3ul gel green 5ul PCR product 4ul 1kb 100 v, 2 hrs.

Non specific amplification in some samples. The IPT1, IPT2 samples will be taken In order to improve amplification, the next PCR I won´t use BSA.

FISH2CB, PCR