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Characterizing Novel Aquatic Phages from the Environment

This study aims to characterize novel aquatic phages from the environment by isolating and characterizing their proteins and DNA. Techniques such as PCR, sequencing, and SDS-PAGE will be used to identify hosts that can be infected and determine protein characteristics. Interesting results from the gel electrophoresis will be verbally described.

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Characterizing Novel Aquatic Phages from the Environment

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  1. Characterizing Novel Aquatic Phages from the Environment 7.391 Minyoung Jang January 19, 2007

  2. 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

  3. SDS-PAGE • Denature isolated, purified proteins by SDS + boiling • Acrylamide gel • Load known samples & molecular weight (MW) markers

  4. 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 Our results

  5. 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.

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