Industry Perspective:DNA Melt Curves

Tami Shinkawa

October 12, 2007

Overview

• 20.309 vs. Industry

• Industry Experiments– Stability Study– Tm Peak Detail Study

• Real Time PCR Systems

• Cool Conclusions

Melt Curve Analysis

• 20.309 Graph– 5 samples– Tm?

• Industry Graph– 96 samples– Tm?

Industry Study: 12 Hour Stability

Industry Study: Tm Peaks Detail

• Tm = 73.48 oC• Step Size = 0.1o

• 2 distinct peaks• Time = 206 min• 96 well plate

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

Industry Study: Tm Peaks Detail

• Tm = 72.66 oC• Step Size = 0.5o

• 1 peak– 2 peaks?

• Time = 40 min• 96 well plate

Real-Time PCR Systems

• Incorporate Melt Curve Technology

• Thermal Cycling vs Real-Time PCR

Cool Conclusions

• Taqman

• Power SYBR vs. Fast SYBR

• Forensics, Fish, Pharoahs

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

? Questions ?