PPT-DNA Sequencing

DNA sequencing How we obtain the sequence of nucleotides of a species ACGTGACTGAGGACCGTG CGACTGAGACTGACTGGGT CTAGCTAGACTACGTTTTA TATATATATACGTCGTCGT ACTGATGACTAGATTACAG ACTGATTTAGATACCTGAC. Learning Objectives. Recap how DNA probes and DNA hybridisation is used to locate specific genes.. Learn how the exact order of nucleotides on a strand of DNA can be determined.. Learn how restriction mapping can be used to determine nucleotide sequences.. MOLECULAR BIOLOGY TECHNIQUES II.. Polymerase Chain Reacton – PCR. DNA sequencing. Amplification of specific DNA fragments. MOLECULAR BIOLOGY – PCR. Cloning and/ or isolation from a genomic library . Dan . Russell. Overview. Prologue: Assembly and Finishing. The Past: Sanger. The Present: Next-Gen (454, . Illumina. , …). The Future: ? (. Nanopore. , . MinION. , Single-molecule). Overview. Prologue: Assembly and Finishing. Craig A. . Praul. Co- Director . Genomics Core Facility. Huck Institutes of the Life Sciences. Penn State University. A very short history of DNA sequencing. I started from the conviction that, if different DNA species exhibited . INTRODUCTION . The most commonly used technology until a few years ago – BAC. WHOLE GENOME SEQUENCING. ADVANTAGES OF WGS. Utility of next – gen sequence reads . The next-generation platforms are effecting a complete paradigm shift, not only in the organization of large-scale data production, but also in the downstream bioinformatics, IT, and LIMS support required for high data utility and correct interpretation.. Lenka Veselovská. Laboratory of Developmental Biology and Genomics . Next Generation Sequencing (NGS) . M. odern high-throughput DNA sequencing technologies. parallel, rapid . Decreasing price, time, workflow complexity, error rate. - . INTRODUCTION. - SANGER DIDEOXY METHOD. - AUTOMATED SEQUENCING. - NEXT. GENERATION OF SEQUENCING METHODS. MISS NUR SHALENA SOFIAN. INTRODUCTION. 1977:. . Frederick Sanger along with Allan . Maxam.  . Nearly . all modern DNA sequencing procedures require a concentrated amount of . single-stranded . DNA known as a . template. . A template is simply a piece of DNA of . sufficient . length and quality to allow for its sequencing (most sequencing methods . Goals:. Review central dogma and limits of DNA. Understand history and recent advances in sequencing. Understand the process (sequencing by synthesis) used to generate data in this module. What is DNA?. Hardison. Genomics 3_1. 1. 1/20/14. nucleo. s. ides. A nucleo. t. ide has one or more phosphates attached to the 5’ hydroxyl of the (deoxy)ribose. Building blocks of DNA. 1/20/14. 2. Structure of a dinucleotide. Dr. . Sudha. . Kumari. Assistant Professor. Department of Veterinary Microbiology. Bihar Animal Sciences University, Patna. INTRODUCTION . The . DNA Replication Review. Enzymes open . double stranded DNA. Origin of replication. Replication fork. DNA replication in 5’ to 3’ direction. Leading and Lagging Strands. RNA primers. DNA polymerase. DNA Cloning. 3. Introduction . into. host cell. Restriction Enzymes. PCR. Polymerase. Chain. Reaction. Antibiotic Selection. Antibiotics, such as penicillin and ampicillin, kill bacteria. Plasmids can carry genes that give host bacterium a resistance against antibiotics. Figure 8.01. Sequencing—Fragments of All Possible Lengths. During chain termination or . Sanger sequencing. , the target DNA fragments are copied millions of times, but each copy ends at a different nucleotide position. These subsets of fragments end with a fluorescently-labeled nucleotide that reveal the identity of the final base. The final sequencing data are a series of fluorescent peaks that correspond to the original template .