Protocol for SLST for P. acnes

Original primer set
Forward primer
5'-CAGCGGCGCTGCTAAGAACTT-3'
Reverse primer
5'-CCGGCTGGCAAATGAGGCAT-3'

New primer set
Forward primer
5’-CGCCATCAAGGCACCAACAA-3’
Reverse primer
5’-ATATCGGCCCGTATTTGGGC-3’

PCR reaction mixture
PCR reactions are carried out in a 25 µl volume containing:
10 µl 5’-PRIME Hotmastermix (5 PRIME, Hamburg, Germany)
3 µl of each primer (10 µM)
4 µl of PCR grade H2O
5 µl of diluted DNA

PCR cycling conditions
Start with an initial denaturation at 96°C for 40 seconds,
followed by 35 cycles at:
[94°C for 35 seconds]
[55°C for 40 seconds]
[72°C for 40 seconds]
Finally extend at 72°C for 7 minutes.

Reference
Scholz CFP, Jensen A, Lomholt HB, Brüggemann H, Kilian M (2014) A Novel High-Resolution Single Locus Sequence Typing Scheme for Mixed Populations of Propionibacterium acnes In Vivo. PLoS ONE 9(8): e104199. doi:10.1371/journal.pone.0104199
Go to PLoS ONE