2020-03-06-16SFD-2Rdeg-p152
16SF/D, 16S2R-degenerate Primer (Fish-16S), amplify 200 pb, Deagle et.al.2007
PCR #2, Pag 152, 03 06 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 1).