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01_Mapping_and_sequencing

01_Mapping_and_sequencing - MCB182 Class Notes Quick Molbio...

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Central dogma of molecular biology : The information flow is generally from DNA to RNA to protein. From an -omics perspective, we would say genome to transcriptome to proteome (to metabolome). There are a lot of other -omes we could add. Nucleic acids : Visit the Wikipedia page on DNA . Most of this should be review for you. You do not need to memorize the chemical structures, but should be able to interpret/draw a cartoon of DNA showing the ring structures and hydrogen bonds between base pairs. You ought to be able to recognize the difference between DNA and RNA (the 2’ OH). You should memorize the IUPAC ambiguity codes for nucleotides. The fundamental property of DNA is that it can be denatured (melted) and hybridized (annealed). The melting temperature ( Tm ) depends on length (longer is harder to melt), GC content (GC pairs have 3 bonds), mismatches (reduces Tm), salt concentration (raises Tm), and supercoiling (positive supercoiling raises Tm, negative reduces Tm). Obtaining DNA : DNA can be extracted from an organism, synthesized as oligonucleotides (aka oligos or primers ) or polymerized from a template. The template may be DNA or RNA. In the case of an RNA template, the polymerase is called reverse transcriptase (RT) . This is one of the more useful enzymes for studying the transcriptome because it lets you turn RNA into DNA. The polymerase chain reaction ( PCR ) is one of the most important molecular biology inventions. PCR allows one to amplify a specific region of sequence located between two primers. Cloning : The word clone has many meanings. In molbio it means capturing a piece of DNA and growing it in a host organism. Most commonly, the DNA is inserted into plasmid vector and grown in E. coli or S. cereviseae. Plasmid vectors hold up ~10 Kb, while bacterial artificial chromosomes ( BAC ) hold ~300 kb and yeast artificial chromosomes ( YAC ) hold ~600. MCB182 Class Notes: Quick Molbio Review, Mapping and Sequencing 1/3 Letter Meaning R A or G Y C or T M A or C K G or T W A or T S G or C B not A D not C H not G V not T N all Bold text terms are likely to appear in exams
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Genetic maps : The goal of genetic mapping is to place genetic markers on chromosomes. When creating a genetic map, the first step is to determine which markers are on which chromosomes and then to determine the order and relative distance between markers. Historically,
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