Fast PCR is both
specific and sensitive. QuanTyper™-48 is
able to detect a single gene copy in less
than 14 minutes. Slower ramping in a conventional
block cycler is less sensitive and produces
more non-specific amplification.
The figures below show
the result of a parallel comparison of amplification
efficacy for identical 20 µl samples run
on a QuanTyper-48 vs. ABI 7500. PCR reaction
mixes contained SYBR Green I for real-time
detection. Melting point and agarose gel
analysis was used to discriminate between
specific and non-specific amplification.
The PCR template was human DNA prepared
from blood and 10-fold serially diluted
from 50 ng down to 5 pg (5 pg roughly corresponds
to one human haploid copy of DNA). Amplicon
length is 115 bp. PCR cycling parameters:
98°C for 10 seconds followed by 40 cycles
of [98°C for 0 s; 58°C for 3 s; 72°C
for 3* s].
* 32 s in ABI 7500, which is the shortest
setting allowed by the instrument.
