Comparative genomics of European avian pathogenic E. Coli (APEC)

BMC Genomics

Table 1 8plex, 5plex and 9plex primers and respective cycling conditions. The gene name, primer sequence and amplicon size are reported. Lowercase letters in the gene sequences (8plex) represent the tails added in order to obtain the same annealing temperature

8 PLEX
Gene name	Sequence 5′–3′	Amplicon size bp
R1F +	gcgaaaaGAGTAATGTCGGGGCATTCA	551
R1R +	aggccaTTCCTGGCAAGAGAGATAAG	551
R2F +	gcgaGATCGACGCCGGAATTTTTT	1141
R2R +	gcgagaAGCTCCATCATCAAGTGAGA	1141
R3F +	agccaGGCCAAAACACTATCTCTCA	1785
R3R +	agcgccGTGCCTAGTTTATACTTGAA	1785
R4F +	gcgcgcaTGCCATACTTTATTCATCA	699
R4R +	gcgcTGGAATGATGTGGCGTTTAT	699
K12F +	gcaagTTCGCCATTTCGTGCTACTT	916
k12R +	acgcgcTAATCATAATTGGAATGCTGC	916
chuAF +	aaatttgGACGAACCAACGGTCAGGAT	279
chuAR +	atttagTGTGAAGTGTCAGGAGACGCTG	279
yjaAF +	aaaaaaCCGCCAGTACCAGGGACA	211
yjaAR +	gcagaaaaATGGAGAATGCGTTCCTCAA	211
TSPEF+	gcgaaaaaGAGTAATGTCGGGGCATTCA	152
TSPER+	aaggCGCGCCAACAAAGTATTACG	152

Cycling conditions: 95 °C for 5 min (Initial denaturation), followed by 2 cycles 95 °C (denaturation) for 30s, 50 °C for 30s (annealing), and 72 °C for 60s followed by 33 cycles 95 °C for 30s (denaturation), 58 °C for 30 s (annealing), and 72 °C for 60s (polymerization). On completion of 35 cycles the, a final polymerization was performed at 72 °C for 420 s

ISSN: 1471-2164