PPT-Introduction to dna sequencing

Vince Buonaccorsi Juniata College Cost of sequencing httpswwwgenomegovaboutgenomicsfactsheetsSequencingHumanGenomecost Next 2 nd Generation Sequencing How does Illumina sequencing work. 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 . 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 . 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. A I . Bhat. Indian Institute of Spices Research. Calicut. DNA sequencing. Chain termination method . (. Sangers. . et a. l., 1977): In this method, the sequence of a single stranded DNA molecule is determined by enzymatic synthesis of complementary polynucleotide chains, these chains terminating at specific nucleotide positions.. Hardison. Genomics . 3_2. 1. 1/20/14. Second generation sequencing. Michael . Metzker. review. (2010) Nature Reviews Genetics . 11. : 31-46. 1/20/14. 2. Two generations of sequencing technology. Feature. - . INTRODUCTION. - SANGER DIDEOXY METHOD. - AUTOMATED SEQUENCING. - NEXT. GENERATION OF SEQUENCING METHODS. MISS NUR SHALENA SOFIAN. INTRODUCTION. 1977:. . Frederick Sanger along with Allan . Maxam. - . INTRODUCTION. - SANGER DIDEOXY METHOD. - AUTOMATED SEQUENCING. - NEXT. GENERATION OF SEQUENCING METHODS. MISS NUR SHALENA SOFIAN. INTRODUCTION. 1977:. . Frederick Sanger along with Allan . Maxam. How we obtain the sequence of nucleotides of a species. …ACGTGACTGAGGACCGTG. CGACTGAGACTGACTGGGT. CTAGCTAGACTACGTTTTA. TATATATATACGTCGTCGT. ACTGATGACTAGATTACAG. ACTGATTTAGATACCTGAC. TGATTTTAAAAAAATATT…. DNA polymerase (copy DNA), restriction endonucleases (cut DNA), ligases (join DNA). DNA cloning – vector (plasmid, BAC), PCR. genome mapping. relative locations of genes are established by following inheritance patterns. David . Tse. . Dept. of EECS. U.C. Berkeley. LIDS Student Conference. MIT. Feb. 2, 2012. Research supported by NSF Center for Science of Information.. TexPoint fonts used in EMF: . A. A. A. A. A. A. 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. 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. First, anneal the primer to the DNA template (must be single stranded):Then split the sample into four aliquots including the following nucleotides: When a DNA polymerase (e.g. Klenow fragment) ispro 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 .