Pitt BIOSC 1850 - Exam 3 Study Guide (13 pages)

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Exam 3 Study Guide

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Exam 3 Study Guide


Study Guide
University of Pittsburgh
Biosc 1850 - Microbiology
Microbiology Documents

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BIOSC 1850 1st Edition Exam 3 Study Guide Textbook Chapters 10 11 13 14 15 Chapter 10 Microbial Genomics 10 1 Genome Sequencing Sanger Dideoxy Sequencing cloning of the fragment to be sequenced DNA synthesis gel electrophoresis o Primer is added complementary to flanking vector sequences o Bases are attached to a tri phosphate group for marking and are added to the mix These altered bases stop further replication so we re left with a lot of shorter bases o Gel electrophoresis is run for each of the four bases to determine sequence Primer Walking consecutive runs of sequencing and primer for longer sequences Next Generation Sequencing Methods not reliant on gel electrophoresis o Pyrosequencing the four altered bases are added one at a time If it matches it produces light due to the release of PPi which gets converted into ATP that drives a light producing reaction o 454 DNA Sequencing DNA fragments don t have to be added into vectors Shotgun Sequencing shearing DNA into short pieces sequencing those fragments Computer programs are used to map the overlapping sequences to link together individual sequences 10 2 Genomic Analysis of Gene Expression Genomic library a collection of cloned DNA fragments that comprise the entire genome of that organism cDNA Library genomic library of the mRNA sequences only o Varies under different conditions Proteome collection of proteins present in a cell under certain conditions Transcriptome set of transcripts encoded by each gene within a genome o Transcription patters o Northern Blotting Binding a probe to a known sequences of mRNA to detect its presence and the level of transcription o Microarray Simultaneous examination of the transcriptional activity of all genes in the genome Relative abundance of each transcript determined by the color output 10 3 Comparative Genomics Determining evolutionary relationships Paralogs genes that arise from duplication events Orthologs genes with the same function in different organisms that have

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