S-STEM Scholar Carime Flores' Blog

  Introduction  The following week, I made some media and did gram-staining on D. caeni and D. sonorensis. My classmates Emme and Bryan did a presentation discussing different chemical tests and their purposes. The plan is to use some chemical tests to characterize Deinococcus Sonorensis. We also had a discussion where we learned the difference between Rt-PCR, q-PCR, and Rt-qPCR.  Methods For the gram staining, I began by getting the bacteria (sonorensis and Caeni) on two separate microscope slides. First, you turn on the flame and sterilize the loop. Then after letting the loop cool, slightly lift the lid of your plate and scoop some of the bacteria with the loop. After spreading the bacteria on the slide finally heat fix it by passing the slide over the flame. Next, you have to complete the steps of the gram stain. You add a couple of drops of crystal violet on top of the bacteria, and after a minute you wash the slide with deionized water. Repeat this step with iodine....

  Introduction During this week we learned about the fluorescent microscope and the dyes that are used for it. On Friday we prepped and did a q-pcr for our E. Coli, along with starting sugar fermentation biochemical tests by inoculating some tubes.  Method The biochemical test that was performed was to check if D. Sonorensis can ferment certain sugars and in this case, it was Sucrose, fructose, and glucose. The broth is a reddish color. If the sugars are fermented, acids will be produced and will change the pH below 7 of the broth therefore changing the color to yellow. If the sugars are not fermented the bacteria will consume proteins and produce alkaline change the base above 7 changing the media a dark red. On Thursday 6 tubes were inoculated while three were left without inoculation as a negative control. Three of the tubes were inoculated with staph as a positive control. The last three tubes were inoculated with  D. sonorensis.    Steps of inoculation plat...

  Introduction This Week I learned the recipe for different media. I Helped make different media. Along with making media, we practiced some conversation math. We learned and practiced the C1 V1 = C2 V2 equation. We got 40 sample questions and had to solve them.  Methods the general step of making media  -Start with half of the whole amount of water (choose a flask that holds the double volume of liquid so when it's autoclaved it won't spill over)  -weight out your water with a graduated cylinder  -prep the scale by zeroing it and then grab a paper towel, then grab your dry ingredients -Weigh out each ingredient then add it to the flask with water - mix the mixture to dissolve all the powder ingredients (clean your scope of each different ingredient with your paper towel)  (Once you take out some of the ingredients from its container you have to throw them out because you can't put them back since it could contaminate the ingredients)  -add in the rema...

  Introduction This week was the biotech boot camp where we had a basic introduction to lab safety. We learned about the protocol for entering, leaving, and being in the lab. When you enter the lab you leave any drinks in the red bin (we are not allowed to drink anything), then you wash your hands and sign into the lab book. You also need to clean your table with fantastic. While being in the lab you should keep your space clean, be mindful of others, wear your goggles, and write in your lab notebook. When you’re leaving the lab you should clean and put everything away in your area. Write your final note in your lab notebook about what you did and plan for next time. Then wash your hands and sign out of the lab book. We learned some general rules like giving Chad and Jonathan 2 or 3 days in advance when you will need something. Along with always asking questions when you are unsure, never opening anything without consulting someone about it, and doing all your research outside of t...

  Introduction The previous week, I researched different chemical tests that help characterize bacteria. These tests range from the bacteria's physical features to reactions to growing in different media. We also had a discussion about the Amc cycle. The discussion was a surface explanation of the cycle. I also learned how to make double TGY  and R2A media. I also learn the steps of RNA isolation. the following Friday, I assisted Madison as we attended to an RNA isolation of E. coli.  Methods  Characteristic tests First, you can assess with your eyes anything particular about the appearance of the bacteria, the way the colonies look, its edges, and things you think are worth mentioning. There is the gram-stain test to figure out if a bacteria is gram-negative or gram-positive. after the stain, you can look at the bacteria under the microscope to figure out the bacteria's arrangement and morphology. You can try to grow them in selective and differential media, these m...

  Introduction  This previous week, I helped with an ethanol precipitation and made some media. Ethanol precipitation from what I understand is a process used to “draw out” (precipitate) DNA or in our case RNA. The results were not what we hoped and ultimately had to begin from step one where we needed to innoculate some tgy plates with D. caeni. We made 250 ml of Tgy and LB.  Methods and results We started the ethanol precipitation on Monday the 5th and added 150 µl RNA into 6 tubes. Then we proceeded to add 500µl of ethanol, along with 20µl sodium acetate. The last thing we added was 2 µl of glycogen blue to be able to see the pellets the next day. We left the tubes in -80 deep freeze overnight. Then on Tuesday, we put the tubes in the centrifuge at full speed for 15 minutes. While this process took place we made 250 ml of Tgy and LB. we measured out the ingredients Tgy LB -Deionized water 250mL           ...