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1. Thal University Bhakkar Presentation Submitted to : Dr.M.Kabir Submitted by : Aqsa Zulfiqar Roll No : BOTN51BF20R049 Class : BS Botany (7th) Subject : Ecology II Topic : DNA Sequencing 2024/1/5 1 DNA Sequencing

2. Contents : Definition History Ingredients Methods Steps Application 2024/1/5 2 DNA Sequencing

3. Definition: DNA Sequencing is the process of determining the nucleic acid sequence (the order of nucleotides in DNA). It includes any method which is used to determine order of the four bases : Adenine, Guanine , Cytosine,Thymine. ( A, G, C, T ) 2024/1/5 3 DNA Sequencing

4. History: 1870--Friedreich Miescher:" discovers DNA " 1940--Avery: proposes DNA as "Genetics material " 1953--Watson &crick:( double helical structure ) 1977--Sanger : Dideoxy ChainTermination 1977--Gilbert: Chemical Degradation 1986-- Partial automation 1990-- Cycle sequencing ,improved sequencing enzymes, improved flourscent detection schemes. 2 002-- NGs: 454 Pyrosequencing 2024/1/5 4 DNA Sequencing

5. Ingredients: Primer DNA Polymerase DNA Nucleotides (dATP,dGTP,dTTP,dCTP) 2024/1/5 5 DNA Sequencing

6. Methods: To determine the order of the nucleotides bases adenine, guanine cytosine, thymine in a DNA molecule two methods were used : Sanger sequencing ;It was developed by Friedrick Sanger and his colleagues in 1977. It is also called Chain Termination Sequencing Method. Maxam and Gilbert: Maxam &Gilbert sequencing is a method of DNA sequencing that developed by Allan Maxam & Walter Gilbert in197---1977. 2024/1/5 6 DNA Sequencing

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8. Steps: There are three main steps to Sanger sequencing : DNA sequence for chain termination PCR.The DNA sequence of interest is used as a template for a sspecial type of PCR called Chain Termination PCR . Size separation by gel electrophoresis . Gel analysis & determination of DNA sequence. 2024/1/5 8 DNA Sequencing

9. Application : Forensic :to help identify individuals because each individual has a different genetic sequence. Medicine : can be used to help detect the Genes which are linked to various genetic disorder such as muscular dystrophy. Agriculture :The mapping and sequencing of a genome of microorganisms has helped to make them useful for crops and food plants. 2024/1/5 9 DNA Sequencing

10. Reference: https://www.sigmaaldrich.com/PK/en/technical-documents/protocol/genomics/sequencing/sanger-sequencing. https://www.slideshare.net/mallappashalavadi/dna-sequencing-18897579 https://www.cd-genomics.com/blog/dna-sequencing-definition-methods-and-applications/ 2024/1/5 10 DNA Sequencing