PPT-DNA sequencing I

Historical method Sanger N chain termination Latest method ion torrent seq via pH measurement Both rely on DNA polymerase to copy template ie sequencing by synthesis. DNA sequencing. How we obtain the sequence of nucleotides of a species. …ACGTGACTGAGGACCGTG. CGACTGAGACTGACTGGGT. CTAGCTAGACTACGTTTTA. TATATATATACGTCGTCGT. ACTGATGACTAGATTACAG. ACTGATTTAGATACCTGAC. 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 . Sbi4up. Mrs. franklin. DNA Sequencing. DNA sequencing is a method commonly used by scientists to determine the nucleotide sequence for a particular gene. The Sanger sequencing method enables Frederick Sanger to determine the entire genome of a bacteriophage (5386 . 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 . 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.. How we obtain the sequence of nucleotides of a species. …ACGTGACTGAGGACCGTG. CGACTGAGACTGACTGGGT. CTAGCTAGACTACGTTTTA. TATATATATACGTCGTCGT. ACTGATGACTAGATTACAG. ACTGATTTAGATACCTGAC. TGATTTTAAAAAAATATT…. Knowing how many genes determine a phenotype (Mendelian and/or QTL analysis), and where the genes are located (linkage mapping) is a first step in understanding the genetic basis of a phenotype . A . 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 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. 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 . The Sanger method. DNA sequencing is the primary method for gene and protein characterization. . (protein sequence can be predicted from DNA sequence). . https://. www.illumina.com. /techniques/sequencing/. the DNA sequencing?. DNA sequencing determines the order of the four chemical building blocks - called "bases" - that make up the DNA molecule. . The . sequence tells scientists the kind of genetic information that is carried in a particular DNA segment.