Question:
How do the Taqman probe and SYBR green fluorophore work in qPCR?
Author: Mirte SchattorieAnswer:
Taqman probe: under normal conditions, the probe is coiled up, meaning that fluorescein is near the quencher of the probe, limiting the fluorescent signal. when the target sequence is found, the probe will uncoil and bind to the sequence. as new DNA is created, the probe starts being degraded, which means that fluorescein is separated from the quencher and a fluorescent signal is shown. this is very specific. SYBR green: SYBR green dye binds to the minor groove of the DNA non-specifically and emits fluorescence. this provides information on each cycle of amplification and about the melting temperature.
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