16SF/D, 16S2R-degenerate Primer (Fish-16S), amplify 200 pb, Deagle et.al.2007

PCR #3, Pag 154, 03 10 2020

PCR reaction

• ~ 30 ng/ul DNA
• 0.5 ul primer (10um)
• 6.25 ul Hoststart Qiagen
• 12.5 ul total reaction
• 0.5 ul BSA

Cycling conditions (Thomsen et al. 2012)

• 10 min to 94°C
• 35 cycles: 94°C 30 s, 54°C 30 s, 72°C 45 s,
• 2 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.

Pcr’s products amplified very well. I will use it for the sequencing test (Replicate 2).

, PCR