added template DNA) enables to check whether or not the reaction mixture was
contaminated. All the raw fluorescent results are later analyzed using software
connected to the instrument.
There are two basic approaches to the interpretation of the output data. First, the
absolute quantification gives the exact number of the targeted DNA molecules by
comparison with DNA standards using a calibration curve. On the other hand,
Table 34.1 Example of specific primers for target genes. Adapted from Dolinová et al. (2017)
Target
gene
Target microorganism
Primer Primer sequence (5
0 -3
0 )
pceA
Dehalospirillum
multivorans
Primer
F
Primer
R
GCTGCTGGATGGACCTTAGA
CATAGCGATACCTGCAACGA
Dehalobacter spp.;
Dehalococcoides
mccartyi;
Desulfitobacterium
spp.; Desulfomonile
spp.; Desulfuromonas
spp.
Primer
F
Primer
R
ACCGAAACCAGTTACGAACG
GACTATTGTTGCCGGCACTT
tceA
Dehalobacter spp.;
Dehalococcoides
mccartyi;
Desulfitobacterium
spp.; Desulfuromonas
spp.; Geobacter spp.;
Sulfurospirillum spp.;
Primer
F
Primer
R
ACGCCAAAGTGCGAAAAGC
TAATCTATTCCATCCTTTCTC
vcrA
Dehalobacter spp.;
Dehalococcoides
mccartyi;
Desulfitobacterium
spp.; Desulfuromonas
spp.; Geobacter spp.;
Sulfurospirillum spp.
Primer
F
Primer
R
TGCTGGTGGCGTTGGTGCTCT
TGCCCGTCAAAAGTGGTAAAG
bvcA
Dehalobacter spp.;
Dehalococcoides
mccartyi;
Desulfitobacterium
spp.; Desulfuromonas
spp.; Geobacter spp.;
Sulfurospirillum spp.;
Primer
F
Primer
R
TGCCTCAAGTACAGGTGGT
ATTGTGGAGGACCTACCT
etnC
VC-assimilating
bacteria
Primer
F
Primer
R
CAGGAGTCSCTKGACCGTCA
CARACCGCCGTAKGACTTTGT
etnE
VC-assimilating
bacteria
Primer
F
Primer
R
AACTACCCSAAYCCSCGCTGGTACGAC
GTCGGCAGTTTCGGTGATCGTGCTCTTGAC
VC vinyl chloride
652
A. Ševců et al.