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Characterizing Novel Aquatic Phages from the Environment 7.391 Minyoung Jang January 19, 2007 Overview • Take water sample from the environment • Isolate: column, extract protein (CsCl), DNA (phenol chloroform) • Characterize: titer, identify hosts that can be infected, PCR, sequencing, digests SDS-PAGE • Denature isolated, purified proteins by SDS + boiling • Acrylamide gel • Load known samples & molecular weight (MW) markers Our results • Gel image here– LARGE, easy to see • Label all lanes • Label MW markers • Label important bands • Describe conditions: time run, voltage, & anything non-standard, how much you loaded • ALL LABELS SHOULD BE EASY TO SEE! • Describe interesting results VERBALLY (not written) – How many different distinct bands do you see? Any particularly strong? What weight? – Do any bands overlap with your “host cell” sample bands (ie those bands are not from phage) – Etc. – Be ready to answer questions about your protocol & data Bioinformatics Homework • • • • • • • Protein identified Source organism Protein function Amino Acid length Known structures, family of proteins Other functions of homologs LIST ALL SOURCES!! (can footnote it & abbreviate [1] [1] Weigele, et al., 2007. Journ. Name Abbrev. Form.
