SYBR green and TAQ man Flashcards
What is the main difference between SYBR Green qPCR and TaqMan PCR?
SYBR Green qPCR uses SYBR Green dye to detect double-stranded DNA
TaqMan PCR uses a TaqMan probe to detect amplicon formation.
Why is SYBR Green qPCR considered more cost-effective compared to TaqMan PCR?
SYBR green only needs SYBR green dye
TAQ man needs a probe which is more expensive
How does SYBR Green dye work in qPCR?
SYBR Green dye fluoresces when it is put into double-stranded DNA, which lets the fluorescence intensity as amplicon is produced be monitored
What is the function of a TaqMan probe in TaqMan PCR?
The TaqMan probe is a single-strand oligonucleotide that binds to a complementary sequence between the forward and reverse primers.
How does the TaqMan probe contribute to fluorescence detection in TaqMan PCR?
the fluorescence emitted by the reporter is continually absorbed by the quencher as long as the probe remains intact. However, as amplicon is produced, the polymerase degrades the probe, separating the reporter from the quencher and allowing fluorescence detection.
Why does the fluorescence signal increase in TaqMan PCR as more amplicon is produced?
As more amplicon is produced, more probes bind to them and are subsequently destroyed in each PCR cycle, leading to an increase in fluorescence signal.
Which part of the TaqMan probe breaks away first during PCR amplification?
The nucleotide at the 5’ end of the TaqMan probe, which contains the attached reporter, breaks away first due to polymerase activity.
What happens to the fluorescence signal when the reporter and quencher are separated?
the fluorescence signal is detected by the instrument because the quencher can no longer absorb the emitted fluorescence.
