实时荧光定量PCR应用定量.PPTVIP

* Classic PCR – Strait forward. Red triangles are the blocking agent for the polymerase (my skills with powerpoint are limited) ? * SYBR Green I dye has an increased fluorescent intensity when bound to double stranded DNA. * When sample is denatured, signal drops. When more double stranded DNA is generated, the signal increases proportionally with the amount of dsDNA present. * * How does melt curve work? – After the main amplification run, usually 40 cycles, We set the instrument to collect periodic fluorescent readings of our samples as we slowly ramp the block from 55 to 95 degrees ce