MBIO 2420 Lecture Notes - Lecture 4: Sybr Green I, Taqman, Molecular Beacon
7 Dec 2016
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Multiplexing - can analyse different genes at same time in different samples. Lux - made by invitrogen: all can except sybr green, major disadvantage of sybr green. It will analyse the melting point of your product: all except taqman and lightcyder. Zero chance of false positive: and a third, a probe that has a fluorophore (5") and quencher (3") Quencher absorbs any light fluorophore emits: so whenever you try to excite the sample, the energy goes from fluorophore to quencher and no light is emitted. The polymerase does its job and cuts off the fluorophore and it goes into the buffer: so now when you excite the fluorophore the quencher does not absorb the light. Similar to taqman, with fluorophore and quencher: on the hairpin, fluorophore and quencher are literally right by side, the probe is complementary to a part on the dna. When they bind to a sequence the hairpin opens and the fluorophore and quencher are farther apart.
