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DNA Sequencing

The idea of DNA sequencing is to work out the order of nucleotide bases in DNA. This was first done by Sanger et al. who sequenced 500-800 bases at a time and won the Nobel prize in 1980. The Human Genome Project was established in 1990 and was completed quicker than expected due to automation of sequencing techniques.The DNA to be sequenced is mixed with primers, nucleotides, DNA polymerase and terminator bases. Terminator bases will stop DNA synthesis and have coloured fluorescent tags for identification. The mixture is placed in a thermal cycler. At 96°C the strands separate, at 50°C the primers anneal and at 60°C DNA polymerase adds nucleotides to the primers. Each time a terminator base is incorporated the chain terminates so after many cycles DNA of all possible lengths is obtained.

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Specification: OCR Biology A (H420), A-level, 6.1.3 Manipulating genomes. Topic: Genetic Engineering and Biotechnology.

Key terms in this lesson

primer
A short single strand of DNA that binds to a complementary template strand and provides a starting point for DNA polymerase.

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