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Lecture 2[1]

Lecture 2[1]

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Published by MuhammadFakhriAimi

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Published by: MuhammadFakhriAimi on Nov 10, 2011
Copyright:Attribution Non-commercial


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At the end of this lecture, students will be ableto:
define sequencing
Compare between the enzymatic and chemicalsequencing method
Explain how chemical sequencing method iscarried out
Describe the differences of automatedsequencing method compared to the
conventional sanger’s dideoxy method
Lists applications of sequencing
Learning Outcomes:
Sequencing of DNA
DNA sequencing is the determination of the precisesequence of nucleotides in a sample of DNA
Sequencing is principally classified by two methods:
Enzymatic sequencing
(or named after its developers 'Sanger-Coulson- Sequencing')
Chemical sequencing method
(or named 'Maxam-Gilbert-Sequencing' after its developers)
Maxam & Gilbert's method (1976)
-chemical cleavage method using doublestranded DNA-involves 2 steps catalytic process
2 chemicals that selectively attackspurines or pirimidines (step 1)
Piperidine will catalyze phosphodiesterbond cleavage where the base has beendisplaced (step 2)

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