120 likes | 456 Vues
DEVELOPING FLUORESCENT LIFETIME PROBES USING BODIPY DYES. Thomas P. Gonnella. Bodipy TR-X . Bodipy TMR-X . Bodipy 576-X . Probes. RT-PCR. SNP. Dye Labeled Probe. G. G. G. G. G. G. T. T. C. C. C. A. A. A. A. Targets - Extended. WAVELENGTH TIME MATRIX (WTM) of Blank.
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DEVELOPING FLUORESCENT LIFETIME PROBES USING BODIPY DYES Thomas P. Gonnella
Bodipy TR-X Bodipy TMR-X Bodipy 576-X Probes
RT-PCR SNP Dye Labeled Probe G G G G G G T T C C C A A A A Targets - Extended
WAVELENGTH TIME MATRIX (WTM) of Blank WAVELENGTH TIME MATRIX (WTM) of Blank 0.020 0.015 Intensity (mV) Intensity 0.010 (mV) 0.005 0.000 660 l (nm) 10 l (nm) 20 30 585 40 50 60 Time (ns) Time (ns) WTM
Resolved Temporal Components TR-Blank Sample Tau 1: 4.893 Intensity (mV) Tau 2: 1.693 Time (ns) Lifetime
Resolved Spectral Components in TR-Blank Sample % Component 1 : 87.38 % Component 2 : 12.67 l (nm) Lifetime
<t> = 1Iiti2 1Iiti = I1ti2 +I2t22 +… I1ti + I2t2 +... Calculating Average Lifetime
Resistanceto Photobleaching for Hybrodized Products Percent FluorophoresIntact # of WTM’s Collected (avg 5000) Photodecompostion(Blank – Wild – Mutant)
VRN1-sequence: 1488 bp GATGAGACGCTTGACAACAGTGTATTGATGGATGTCTGGTCGGTATACACGCACAGCACAGTACCCCTACTCTAGGACTGGCGAGTATCTTTCATTCATTCCAGAAATACGCGGGTCGGCCAAAAGTAGAAAAATACACTGCGCCCACTCAATCCACGCAGCGCACTGCACTGCACAGCAACGCTTCATGTCAAAAGTCGAGCTCAAGCATGCACGCGATGGACGCGGCGCGAATGACCCGGGCGGCACGACGCGAGTGCCCGCCGCGCCCGCCCGCCTGCCCCGCAGCCGACCTCTCCCAAACGGGACAAGCGAGACGGCCCAAAACGAGCAAGGAAAGCAGCCTCCTACTGTGGCAGCCCGCCCCCACGACCGTCATCTCGCCTTCCATTCCATTTTCCCTGGACGGACCAGACCCGTCCGAGCCGCCCTGACCTAGCCAGCCAGCATTTCCTCTTTCGTCCCCCGCCGCCGTGACCAAAAAAGCAAAAAAGGAAAAAGGGAAAATGCTAAAGGAAAAAACTCCGCTCTTTCCCTTCTTCTAGGCCTAGGGTACAGTAGAATATTATAAAAGGAAAAATTCTGCTCGTTTTTTGCTCTGTGGTGTGTGTTTGTGGCGAGAGAAAATGATTTGGGGAAAGCAAAATCGGGAGATTCGCACGTACGATCGTTCGACACGTCGACGCCCGGCGGGCCCGTGGTGGGGCATCGTGTGGCTGCAGGACCGCGGGGCCCCGCGGGGCGGGCCGGGCCAATGGGTGCTCGACAGCGGCTATGCTCCAGACCAGCCCGGTATTGCATACCGCGCTCGGGGCCAGATCCCTTTAAAAACCCCTCCCCCCCTGCCGGAACCCTCGTTTTGGCCTGGCCATCCTCCCTCTCCTCCCCTCTCTTCCACCTCACCCAACCACCTGATAGCCATGGCTCCGCCGCCTCGCCTCCGCCTGCGCCAGTCGGAGTAGCCGTCGCGGTCTGCGGGTGTTGGAGGGTAGGGGCGTAGGGTTGGCCCGGTTCTCGAGCGGAGATGGGGCGCGGGAAGGTGCAGCTGAAGCGGATCGAGAACAAGATCAACCGGCAGGTGACCTTCTCCAAGCGCCGCTCGGGGCTTCTCAAGAAGGCGCACGAGATCTCCGTGCTCTGCGACGCCGAGGTCGGCCTCATCATCTTCTCCACCAAGGGAAAGCTCTACGAGTTCTCCACCGAGTCATGGTAAATTAAGCACGCGCTGTCTTTAAATTTGTTCCCCAATACGCCTTCGATTTCGATTTCCTGCGCACCGTTCTGGTCCTGCGAGACGACCCGGCCGACCCCAGGGCCTTCTCCATTTCCGCGCTGCTGTTTGGTTGATTCCGTTTGCCCGCTCGCTGTTTCCATCCGATTCTGCCGTGGCTGCTTGCTCGTTTTTCTTAGAATCGATGGGGGAGCTGGCGTTCCGCGCGGCCGCGATTTCTTGCTATGGGGGGTAGGGCGCGCGATGGGTCGCCGGG