DNA Sequencing- SANGER Method

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