Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.
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Transcript of Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.
![Page 1: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/1.jpg)
Industry Perspective:DNA Melt Curves
Tami Shinkawa
October 12, 2007
![Page 2: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/2.jpg)
Overview
• 20.309 vs. Industry
• Industry Experiments– Stability Study– Tm Peak Detail Study
• Real Time PCR Systems
• Cool Conclusions
![Page 3: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/3.jpg)
Melt Curve Analysis
• 20.309 Graph– 5 samples– Tm?
• Industry Graph– 96 samples– Tm?
![Page 4: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/4.jpg)
Industry Study: 12 Hour Stability
![Page 5: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/5.jpg)
Industry Study: Tm Peaks Detail
• Tm = 73.48 oC• Step Size = 0.1o
• 2 distinct peaks• Time = 206 min• 96 well plate
![Page 6: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/6.jpg)
Industry Study: Tm Peaks Detail
• Tm = 73.18 oC• Step Size = 0.3o
– Default Setting
• 1 wavy peak– 2 peaks
• Time = 70 min• 96 well plate
![Page 7: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/7.jpg)
Industry Study: Tm Peaks Detail
• Tm = 72.66 oC• Step Size = 0.5o
• 1 peak– 2 peaks?
• Time = 40 min• 96 well plate
![Page 8: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/8.jpg)
Real-Time PCR Systems
• Incorporate Melt Curve Technology
• Thermal Cycling vs Real-Time PCR
![Page 9: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/9.jpg)
Cool Conclusions
• Taqman
• Power SYBR vs. Fast SYBR
• Forensics, Fish, Pharoahs
![Page 10: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/10.jpg)
References
• Information:– 20.309 Lab Partner Mike Yee– www.appliedbiosystems.com– http://press.appliedbiosystems.com/corpcomm/
applerapress.nsf/ABIDisplayPress/7AAACA21A7A04E5F88257307006342D8?OpenDocument&type=abi
• Images:– www.appliedbiosystems.com– www.stanford.edu/~jmanning/images_big/hatshepsut.jpg– www.sinaicentral.com – www.fletc.gov
![Page 11: Industry Perspective: DNA Melt Curves Tami Shinkawa October 12, 2007.](https://reader035.fdocuments.in/reader035/viewer/2022081512/56649f275503460f94c3fe0c/html5/thumbnails/11.jpg)
? Questions ?