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-Narayani. NM.Sc Human Genetics

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DNA

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So, how to read the sequence ofnucleotides??

y Biochemical methods

y Need:

y Basic research studying biological processes

y

To correlate changes in gene with a diseased conditiony Forensics

y Diagnostics

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Three main methods

y Maxam-Gilbert sequencing

y Chain termination method

y Dye termination method

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Maxam-Gilbert method

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Sanger Technique

y a.k.a chain terminationmethod

y Use of ddNTPS

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Dye termination method

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Automated DNA sequencing

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Applied Biosystems PRISM 377(Gel, 34-96 lanes)

Applied Biosystems PRISM 3100(Capillary, 16 capillaries)

Applied Biosystems PRISM 3700(Capillary, 96 capillaries)

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Pyrosequencing

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Thank you