Fish2degCBL, Fish2CBR Primer (cytb-Fish), amplify 80 pb, Thomsen et al. 2012

PCR #3, Pag 72, Jul 29 2019

PCR reaction

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

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

The IPT3, IPT4, IPT5 and IPT6 will be taken for test sequencing, GSO samples did not amplify. I will add 0.5ul more from BSA

Fish2deg, PCR