Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Exome sequencing wikipedia , lookup
Zinc finger nuclease wikipedia , lookup
DNA repair protein XRCC4 wikipedia , lookup
Homologous recombination wikipedia , lookup
DNA replication wikipedia , lookup
DNA profiling wikipedia , lookup
DNA nanotechnology wikipedia , lookup
DNA sequencing wikipedia , lookup
DNA polymerase wikipedia , lookup
United Kingdom National DNA Database wikipedia , lookup
Mitochondrial DNA Sequencing DNA from Human Cheek Cells Using PCR General Overview • Day 1: Collect and wash cheek cells • Day 2: PCR(Polymerase Chain Reaction) • Day 3: Gel Electrophoresis and Prepare for sequencing PCR • 1. Denature(95°C): breaks DNA strands apart by separating hydrogen bonds • 2. Anneal(50-65°C): Primer begins adding nucleotides • 3. Extension(72°C): Replicates DNA • 25-50 cycles run to amplify DNA(each doubles the DNA) cycle Primers and Enzymes • Primer is the starting point to start polymerase amplifying the DNA • Taq (Thermus aquaticus): bacteria from hot springs contains enzymes that can withstand the high temperatures of PCR Gel Electrophoresis • Verify if product is at 440 base pairs(bp) Sequencing DNA Final Sequence Printout Interesting Facts • Mitochondrial DNA is from the mother • Father DNA is destroyed during development • Inbreeding causes defects to occur over multiple generations • 5’ reads left to right(normal sequence reading) • 3’ reads right to left • Primers are single stranded DNA • EXOSAP will chew up excess primers and nucleotides. This makes for a cleaner sequencing of DNA