Table 1

Effect of amplicon length on PMA-mediated inhibition of DNA amplification from dead cells in qPCR targeting invA genea
Amplicon Sequence of primers or probe Position Amplicon length (bp) CT value with PMA CT value w/o PMA CT value differenceb
Forward 5′-CGTTTCCTGCGGTACTGTTAATTc 197-219
Probe FAM-CCACGCTCTTTCGMGBNFQd 221-233
A Reverse 5′-ACGACTGGTACTGATGATCGATAATGC 261-238 65 23.81 17.75 6.06
B Reverse 5′-ATTTCACGGCATCGGCTTCAATC 293-270 97 29.96 18.41 11.55
C Reverse 5′-GAATTGCCCGAACGTGGCGATAAAT 315-292 119 33.38 20.54 12.84
D Reverse 5′-TCGCCAATAACGAATTGCCCGAAC 326-303 130 31.52 18.34 13.18
E Reverse 5′-TCGCCAATAACGAATTGCCCGAAC 456-435 260 35.53 21.19 15.44

ainvA gene sequence is from GenBank accession number M90846.

bCT value of untreated dead cells minuses CT value of PMA-treated dead cells.

cThe forward primer is shared by five reverse primers.

dThe probe is shared by five primer pairs.

Li and Chen

Li and Chen BMC Microbiology 2013 13:273   doi:10.1186/1471-2180-13-273

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