pcr & sequencing
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PCR & Sequencing
polymerase chain reaction (PCR)
• produce many copies of a specific target segment of DNA
thermocycler
• dNTPs (free nucleotides)
• Taq polymerase
• buffer• 2 DNA primers
Figure 13.25
3
5
Cycle 1yields 2 molecules
Genomic DNA
Denaturation
Target sequence
3
5
3
5
3
5
Primers
New nucleotides
Annealing
Extension
Cycle 2yields 4 molecules
Cycle 3yields 8 molecules;
2 molecules(in white boxes)
match target sequence
Technique
1
2
3
PCR
http://www.dnalc.org/resources/animations/pcr.html
PCR song: ~2minhttp://www.cnpg.com/video/flatfiles/539/
DNA Sequencing
• Relatively short DNA fragments
• Can be sequenced by the Sanger dideoxy chain-termination method
http://www.dnalc.org/resources/animations/cycseq.html
Human Genome Project 1990-2003
Craig Venter’sshotgun-sequncing
-1000bp per day in 1980s-1000bp per second in 2000-100 billion bp per day in 2010!
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