1. Tucson High School Biotechnology Course Spring 2010

2. What is gel electrophoresis? ------- ++++++ wells DNA (-)

3. Loading Buffer pcr products + STAIN

4. DNA Sequencing

5. DNA Sequencing: Basic Principle Sequence of base pairs?? AACGGATCATTCG polymerase

7. first human genome = ten years @ $1 billion The quest for the $1K human genome 2nd: 10,000 instrument days (30 instruments for 1 year) @ $10 million J. Craig Venter 3rd: 454 pyrosequencing, two months @ $2 million James Watson Nature, Nov 2008 -- “effectively” 4 human genomes (@$100K each) - first African genome (Illumina, UK) - first Asian genome (Beijing Genomics, China) - first cancer genome (Washington University, USA) The $1,000 human genome, coming soon?

8. 454 Pyrosequencing - the generations

9. The “pyro” in pyrosequencing http://www.pyrosequencing.com/

10. Images courtesy of Roche (Technical presentation)

11. Pyrosequencing - emPCR

13. Pyrosequencing - image processing

14. Sequence production (Billions of bases/month) 0.50 0.80 1.00 0.30 0.40 0.20 0.10 0 0 0.70 0.60 0.90 Cost: Cents per base.6 1.0.8.4.2 0 0 1.6 1.4 1.2 2.0 1.8 3.1 1989 1991 1993 1995 1997 1999 2003 2005 2001 Courtesy of Phil Hugenholtz (JGI) Sequencing costs