DNA Sequencing- SANGER Method

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• DNA sequence is amplified- Via cloning the DNA

• Using heat the DNA helix is split into template and complementary strand

• Primer is added to the 5' end of the DNA template

• DNA strands added to four test tubes

• All four of the Free nucleotides are added to solution

• Specially Modified DNA nucleotides are added to test tubes separately

• DNA Polymerase is added to solution

• DNA polymerase binds to primer and works in 5' to 3' direction binding nucleotides

• Until specialised DNA nucleotide is reached and sequence terminates.

• Test tubes added to gel, ELECTROPHORESIS splits the varying sequence lengths

• Sequences move from cathode to anode, because of the negative phosphate backbone.

• Lighter, shorter sequences move further.

• DNA is read from the bottom up, producing complementary strand of DNA

• Amplified

• Heat, Split

• Primer

• Four Test Tubes

• Free Nucleotides

• DNA Modification

• DNA Polymerase

• Binding Nucleotides

• Sequence Terminates

• Electrophoresis

• Negative Phosphate backbone

• Lighter, Shorter sequence

• Bottom Up

• Complementary strand of DNA