Comparative analyses between forensically relevant body
fluids were done with the delta Ct (DCt) method. We selected
human DNA as an endogenous control for relative quantification of
target Lactobacillus DNA. The human DNA quantification was
performed using the human specific primer pair (HSP);
forward primer, 50-TTTTGCAGGATCTACAAGTGGA, and reverse
primer, 50-AAGAGGTCTACATGTCCCCTTG (amplicon size, 207 bp)
[23]. The target of HSP is D17Z1 which is known as a
human specific DNA sequence. Ct values obtained from the LCP and the HSP were named Ct[LCP] and Ct[HSP], respectively;
DCt = Ct[LCP] Ct[HSP]. If a DCt value is small, a high level of target
Lactobacillus DNA relative to human DNA is conjectured. The
amplification efficiency of HSP was verified to near 100% using
serial dilutions of human genomic DNA (TAKARA), and Ct[HSP] > 29
were considered to be below the limit of detection since a Ct[HSP] of
29 represents DNA lower than genome DNA in a cell (6.4 pg)
(Suppl. Fig. 2).
Comparative analyses between forensically relevant body
fluids were done with the delta Ct (DCt) method. We selected
human DNA as an endogenous control for relative quantification of
target Lactobacillus DNA. The human DNA quantification was
performed using the human specific primer pair (HSP);
forward primer, 50-TTTTGCAGGATCTACAAGTGGA, and reverse
primer, 50-AAGAGGTCTACATGTCCCCTTG (amplicon size, 207 bp)
[23]. The target of HSP is D17Z1 which is known as a
human specific DNA sequence. Ct values obtained from the LCP and the HSP were named Ct[LCP] and Ct[HSP], respectively;
DCt = Ct[LCP] Ct[HSP]. If a DCt value is small, a high level of target
Lactobacillus DNA relative to human DNA is conjectured. The
amplification efficiency of HSP was verified to near 100% using
serial dilutions of human genomic DNA (TAKARA), and Ct[HSP] > 29
were considered to be below the limit of detection since a Ct[HSP] of
29 represents DNA lower than genome DNA in a cell (6.4 pg)
(Suppl. Fig. 2).
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