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Table 1 Effect of amplicon length on PMA-mediated inhibition of DNA amplification from dead cells in qPCR targeting invA genea

From: Development of a sensitive and specific qPCR assay in conjunction with propidium monoazide for enhanced detection of live Salmonellaspp. in food

Amplicon Sequence of primers or probe Position Amplicon length (bp) C T value with PMA C T value w/o PMA C T 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
  1. ainvA gene sequence is from GenBank accession number M90846.
  2. bC T value of untreated dead cells minuses C T value of PMA-treated dead cells.
  3. cThe forward primer is shared by five reverse primers.
  4. dThe probe is shared by five primer pairs.