2020-03-04-18S-p148
18S_1F 18S-400R (Eukaryotes; nuclear 18S rDNA gene), amplify 336-423 pb, Pochon et.al.2013
PCR #1, Pag 148, 03 04 2020
PCR reaction
- ~ 40 ng/ul DNA
- 0.5 ul primer (10um)
- 6.25 ul BIOMIX polymerase
- 12.5 ul total reaction
Cycling conditions (Touchdown)
- 3 min to 95°C
- 20 cycles: 94°C 30 s, 62°C 30 s, (decresed by 0.5°C at each cycle) 72°C 1 min,
- 12 cycles: 94°C 30 s, 52°C 30 s 72°C 1 min,
- 7 min to 72°C
- 4°C forever
DNA samples were extracted with Macherey Nagel MN kit on 10 22 2019. Non-specific amplification. I will increase annealing T° and reduce DNA concentration to 25ng.