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Showing posts from May, 2024

Week of 5.28.24 - Cont. D. ficus Transformation

 Tuesday 5.28 Prepped TGY broth for cultures (5x 20ml) Prepped TGY agar for plates (300ml) Discussed experiment and transformation with Dr. Tuohy Experimental Plates, (Predicted Result) TGY, (+) TGY +cmr, (+) Control Plates TGY, (+) TGY +cmr, (-) Wednesday 5.29 Poured TGY plates - 6 Poured tgy +cmr 3ug/mL - 2 Performed Transformation protocol - pRAD1 one uptake into D. ficus

Analysis and Evaluation of Research Paper " The flexibility of UV-inducible mutation in Deinococcus ficus as evidenced by the existence of the imuB–dnaE2 gene cassette and generation of superior feather degrading bacteria"

  The flexibility of UV-inducible mutation in Deinococcus ficus as evidenced by the existence of the imuB–dnaE2 gene cassette and generation of superior feather degrading bacteria Introduction This era of the genome has brought in an unprecedented opportunity to engineer microorganisms for a multitude of diverse biotechnological applications. To accomplish this, the work and research requires rigorous scientific approaches rooted in methodical experimental design and execution.  This paper explores the SOS pathway of TLS polymerase, mutagenesis and the generation of superior keratin breakdown pathways. Where polymerase I-III can stall at UV damage such as thymine dimmers in DNA strands, polymerase IV/V or TLS, Translesion polymerase, is able to move along and repair the damage, however by doing this it also induces mutation, a method of survival to continue the replication cycle of DNA. The team then continues to explore these genes by performing knockouts to deduce that the i...

D. Ficus Transformation Cont. & Contamination Concerns

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 5.7.24          Spent majority of the lab prepping and running gel to confirm the presence of pRAD1 plasmid in plasmid extraction performed the week before hand. Electrophoresis Gel - 30ml @1% with new dye     2ul - Plasmid     8uL - PCR H20     2uL - Dye     6uL - 1Kb Ladder Gel ran @ ~70mA for just under an hour.  This left us with an unusual band amount of 4. Where typically we see 3 bands, a supercoiled plasmid, linear plasmid and circular plasmid, here we see four. Went home to research and worked with team to and decided to check plasmid extraction kit for contamination.  It appears I have a multimer, and have been suggested to linearize my plasmid via restriction site and then run a gel again. Will approach team with this idea next week.  5.8  Plasmid extraction kit contamination results.  No contamination found in reagents.