S-STEM Scholar Carime Flores' Blog

Introduction  This following week I changed groups and joined Anh and Jayce. I learned about their project and some of the major processes they use frequently. These processes are creating competent cells, plasmid extraction, and transformation. We did some inoculations with the transformed cells on agar plates that contained antibiotics. Methods  Competent cells 1.) Pipette 1ml of bacteria culture broth, with a Cell density tube Of 109 into a sterile microcentrifuge 2.) Centrifuge for 1 minute using the Centrifuge at 12,008 ram after which a is pose of the supernatant 3.) Resuspend the pellet in the following mixture. -TGY broth:12901 μι -1m CaCi 2: 51.6 μι -DI water: 335.1 μι -cilcerol 60%: 323 μι  4.) Mix well and place the solution to a 2mL cryovial  5.) Store in -80°c freezer Transformation  1.) aliquot 100 µl Component cells into mircocentrifuge tubes 2.) Add 1μι or < 10 µl plasmid solution to the tube 3.) Place on Ice for 15 min 4.) Place in the incuba...

  Discussion  The following week was after the conference. Which passed with a hitch, I was unable to attend the conference because of work. After the conference, we are now discussing what are our next steps, half of our group will be researching a different project. Where are going to do some more gram-staining on Sono grown on tgy and r2a? Along with looking into our next project. We had a meeting where each group presented their poster and discussed their next stages, if they planned to continue researching their topics, or if they would move on to something new. 

  Introduction  This previous week was the last week before the conference. The week was meant for final edits and printing Final drafts. We experience a major setback due to a miscalculation of the progression of our work for the poster.  Discussion The following week was hard. We put in overtime to finish up the poster. We rearranged the layout of our poster, along with the color scheme from blue/purple to lime green/purple. The method had to be redone, they originally were diagrams that were ultimately changed to a flowchart. We gathered the pictures of the chemical test performed and arranged them on the poster with captions. We also had to do some last-minute gram-stains to get better pictures, which ended up creating a point of contention among the group. We did 8 different gram stains on Sono that grew on tgy and r2a. There was a large discussion because we were noting a variation in gram status depending on what media D. Sonorensis was grown on. when gram-stained ...

  Intro  It is the week before the poster is due and the conference. During the week we are still working on finishing our poster and tying up loose ends. We redid some of our chemical tests for the characterization of D. sonorensis. We redid the sim test, the catalyst test, and some gram stains.  Methods  Gram-stain (steps)  Turn on the bunsen burner, and sterilize the loops Then grab some of your sample with the loop and spread it on the slide  Let it air dry and then pass the slide over the Bunsen burner to heat-fix it Then apply the dyes in this order: Crystal violet -1 minute iodine -1minute Ethanol-quick wash Safranin - 1 minute Then blot dry the slide and look at it under the microscope Catalase test Innoculate a plate with D. sonorensis and then incubate Then in the fume hood add drops of hydrogen peroxide directly onto the bacteria on  the plate  SIM test  (this experiment tests the bacteria for reduced sulfur, production of indole,...