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272 Benčina et al.
B
A
PepC
marker
270bp
230bp
PEPC-1
exon- I
PEPC-1
exon- I
intron
without
with
intron
DNase
reactor
+ - + -
exon- II
PEPC-2
RNase
reactor
+ -
exon- II
PEPC-2
RNA
DNA
DNA impurity
DNA
C
D
RNA
RNA
Reverse
transcription
PCR
Fig. 6. (A) In order to distinguish between RNA and DNA, primers used in PCR
and RT-PCR were chosen to anneal at different exons of DNA, which causes that PCR
fragment of DNA is 40 base pairs larger than RT-PCR product of RNA. (B) Schematic
presentation of either PCR products of DNA or RT-PCR products of RNA and
DNA passed enzyme reactor [deoxyribonuclease (DNase) or ribonuclease (RNase)] or
Convective Interaction Media disk monolithic column of the same chemistry (epoxy
or CDI) but without enzyme. (C) The PCR products of genomic DNA isolated from
Aspergillus niger passed through DNase reactor and control reactor. (D) The RT-PCR
products of total RNA isolated from A. niger passed through DNase or RNase reactor
and control. Simultaneously with samples, PCR or RT-PCR was performed on plasmids
containing PepC gene with and without intron, and presence of PCR products was
determined together with the PCR or RT-PCR products of the sample. Products (10
μl) were loaded on 1.6% agarose gel stained with ethidium bromide. Flow rate was 0.1
ml/min.