DNA Sequencing

Taste Test

• How can you run multiple tests on a small sample size?

Polymerase Chain Reaction

• Replicate DNA regions• Quick process• Occurs within lab– No nucleus required

• 3 main steps– Denaturation– Annealing– Elongation

How many in 5 cycles?

When will you have only target DNA?

Gel Electrophoresis

• Chromatography-like• Uses chemical and physical properties of DNA• Separates fragments of DNA• Negative and positive terminals

Chain Termination Method

• Frederick Sanger– Sometimes referred to as Sanger method

• Labelled dideoxynucleotide (ddNTP)– Used as dyes– Stops elongation process

• Fluoresce under laser light

Sanger Method

Whole-Genome Shotgun Method

• Craig Venter• Faster and longer sequences• DNA randomly cut– Passed through pressurized syringe

• Cloned in plasmids• Sequenced using Sanger method• Computer analyzes partial sequences

Shotgun Sequencing

What is one major drawback?

Bioinformatics

• Analyze sequences• Structural genomics– Locate genes within genome

• Functional genomics– Role proteins play

• Nanopore sequencing– Tiny sub-microscopic holes

• Microarray– Use probe to locate gene

Homework

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