BIO250 Lab 2 FINAL

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“ Lab 2 Culturing & Aseptic Technique BIO250L ” Student Name: Ani Harutyunyan Access Code (located on the lid of your lab kit): AC-Z2CDS9 Lab Report Format Expecta0ons U"lize college level grammar and professional forma4ng when comple"ng this worksheet. Submissions without proper forma4ng, all required photos or suﬃcient responses will be rejected. Pre-lab Ques>ons 1. This lab includes two experiments that each look at different aspects of culturing microorganisms onto growth media. What concepts does each experiment focus on? Why do you think these concepts are important to the field of microbiology? Experiment 1- highlights microbial culture and isolaIon, showcasing the analysis of various microorganisms from surfaces Experiment 2- examines the significance of effecIve hand hygiene in reducing microbial transmission, emphasizing the pracIcal use of asepIc methods 2. In this lab, the experiments will allow or call for the use of four inoculaIon tools. List these tools and provide a scenario in which you would use each tool. ○ CoPon Swabs: Get microbes from surfaces. ○ Sterile CoPon Swabs: Get microbes for the control plate without contaminaIon. ○ InoculaIng Loop: Put microbes on agar plates for isolaIon. ○ Transfer PipePes: Precisely place liquids on agar plates, like in Experiment 4 for yeast. 3. Describe a piece of equipment used in biotech labs to extend the growth paPern of microbes. Focus parIcularly on those used with E. coli used to make human insulin. bioreactor- provides controlled condiIons for microbial culture, including temp., pH, and nutrient supply. 4. You’re a physician trying to isolate bacterial colonies from the human gut in an aPempt to diagnose a gastrointesInal infecIon. You streak your sample on a growth media containing glucose, amino acids, and salts containing both sulfur and phosphorus with a pH of 7 . You incubate the plates in aerobic condi>ons at 37 ˚C for three days, at which point you can see clear bacterial colonies forming on the plate. Would you feel confident in staIng that you had successfully cultured all the bacteria from your gut sample? Why or why not? No, culturing bacteria from a gut sample under aerobic condiIons at 37˚C on a specific medium yields growth of some bacterial types but does not capture the full diversity of the gut microbiome.

“ Lab 2 Culturing & Aseptic Technique BIO250L ” EXPERIMENT 1: AGAR PLATE PREPARATION AND BACTERIAL INOCULATION Introduc>on Ques>ons 1. Describe the objecIves of Experiment 1. State the goal of the experiment and the learning outcome that it is trying to convey to you as a microbiology student. Experiment 1 in microbiology equips us as students with fundamental skills in handling microorganisms and understanding growth condiIons, an essenIal foundaIon for advanced studies and prevenIng contaminaIon. 2. Proper asepIc technique is crucial to ensuring the growth of pure bacterial colonies. What will you do in this lab to demonstrate that you are using proper asepIc technique? - Washing hands properly -work place+tool sterilizaIon -avoiding airborne contaminants -ensuring proper applicaIon technique to prevent contaminaIon In a laboratory se_ng, what are three ways you can properly sterilize culturing equipment? ○ Dry heat ○ Autoclaving ○ UV 3. What method of sterilizaIon will you use in this experiment? autoclaving 4. What results do you expect to see with this experiment? For example, do you expect to see growth from all surfaces or just some? Are there parIcular strains you expect from the surfaces you swabbed? bacterial growth on agar plates from swabbed surfaces, but the types and amounts of bacteria will vary depending on the sources I sampled, with some surfaces having more diverse microbial populaIons than others.

“ Lab 2 Culturing & Aseptic Technique BIO250L ” Data and Observa>ons In the table below, report your observed growth corresponding to each plate number and its corresponding source. Then provide a photo of your growth plates. Your data should have a quanItaIve aspect to it, such as a count of the number of colonies that grew. Your data must also correspond to the photo you include for credit. Table 1: Colony Growth Provide a clear, high resoluIon photo of your plates aber incubaIon with the lids removed. For credit on this lab, your photo must show the growth that is reported in your tabulated data and must also include your handwriPen name in the background. Plate Number Source Growth (Color, Amount, Shape, etc.) 1 Shoes yellow, generous amount, circular 2 toilet handle A slightly pale color, a medium amount, and a mostly round shape with slight irregulariIes 3 Air borne dark yellow, medium amount, mostly round but irregular with slight growth around the plate. 4 Hands white, mulIple small, round, and raised colonies 5 Mouth some small and white, others larger with a yellow hue, and the irregular ones.

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“ Lab 2 Culturing & Aseptic Technique BIO250L ” Sample 1 Sample 2 Sample 3 Control Airborne Contaminatio n

“ Lab 2 Culturing & Aseptic Technique BIO250L ” Results and Discussion 1. Describe what morphological traits (i.e., size, shape, arrangement, color, margin, etc.) you observe from the growth on your plates. Be as specific as possible in the characterisIcs. shoes- yellow, generous amount, circular Toilet handle- A slightly pale color, a medium amount, and a mostly round shape with slight irregulariIes Airborne- dark yellow, medium amount, mostly round but irregular with slight growth around the plate. hands- white, mulIple small, round, and raised colonies Mouth- some small and white, others larger with a yellow hue, and the irregular ones 2. Based on the morphological qualiIes you observed on your plates, and using any resources available to you (i.e., a textbook or internet sources such as the CDC website), look up what bacteria you may have found on the surfaces you swabbed. - Staphylococcus species -Bacillus species -Pseudomonas species 3. For the colonies hypothesized in QuesIon 4, are you surprised to find this type of microbe on this surface? no 4. Looking at your control, did you perform proper asepIc technique or were your plates contaminated? If contaminaIon did occur, list the possible sources and how you can prevent contaminaIon in the future. There is a risk of airborne contaminaIon during experimental plate work, and it appears that my plates may have been affected, as the growth on the plate exposed to airborne contaminaIon resembled that of the sterile swab plate. To prevent contaminaIon in the future, I shall maintain strict asepIc techniques and take bePer precauIons to prevent any airborne parIcles from contaminaIng my experiment. 5. Compare the colonies that grew on your swab plates to the Airborne ContaminaIon plate. Did any of your experiment plates grow colonies similar to your Airborne ContaminaIon plate? How would you describe the risk of airborne contaminaIon in your experiment? Observing similariIes between the colonies on my hands and mouth plates and those on the Airborne ContaminaIon plate indicates a potenIal risk of airborne contaminaIon, underscoring the need for more stringent precauIons in future work.

“ Lab 2 Culturing & Aseptic Technique BIO250L ” EXPERIMENT 2: BACTERIAL TRANSFER TO STAB TUBES AND AGAR PLATES Introduc>on Ques>ons 1. Describe the objecIves of Experiment 2. State the goal of the experiment and the learning outcome that it is trying to convey to you as a microbiology student. The objecIve is to acquire knowledge on bacterial transfer methods and the process of inoculaIng bacteria into stab tubes and agar plates. The desired learning result is to comprehend the effecIve transfer of bacterial cultures while avoiding contaminaIon. 2. What results do you expect to see in Experiment 2? I expect the bacterial cultures to be transferred smoothly, leading to disInct bacterial growth on the agar plates and visible growth in the stab tubes. The essence of this experiment is to demonstrate our ability to proficiently manage bacteria and culIvate them in different media.

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“ Lab 2 Culturing & Aseptic Technique BIO250L ” Data and Observa>ons Record your observaIons in the tables below. Table 2: Ini>al Reserved Plate Colony Growth Observa>ons Table 3: Final Plate and Stab Tube Growth Observa>ons Plate Sample Appearance of the original colonies (morphology, etc.) 1 White/circular/cloudy/irregular edges 2 Glossy, semi-transparent/golden-yellow 3 opaque/Large/circular/smooth/yellow Sample Form Growth (Y/N) Same Appearance as Ini>al Plate (Y/N) Successful Transfer? (Y/N) 1 Plate Y N Y 1 Stab Tube Y Y Y 2 Plate Y Y Y 2 Stab Tube Y Y Y 3 Plate Y Y Y 3 Stab Tube Y Y Y Control Plate Y Y Y Control Stab Tube Y Y Y

“ Lab 2 Culturing & Aseptic Technique BIO250L ” Arrange your plates and tubes as shown below. Remove the lid from the plate and place the stab tube next to the matching plate. Then take a high resoluIon photo and insert it below with your handwriPen name in the background. For credit on this lab, the growth that you report in the above table must correspond to the photo below, and the photo must be of a high enough resoluIon to assess the reported data. Plate/Tube #1 Plate/Tube #2 Plate/Tube #3 Control

“ Lab 2 Culturing & Aseptic Technique BIO250L ” 1. Imagine you are performing an experiment that requires the transfer of cells to different plates which will grow bacteria over an extended period of Ime (days or weeks). Describe why an unsuccessful transfer or growth with a different appearance from the iniIal plates would be a problem in this scenario. If the bacteria transferred don't grow as expected or show different characterisIcs from the iniIal samples, it could seriously affect the experiment's validity, leading to wrong conclusions and wasted effort. Consistent bacterial growth and characterisIcs are essenIal for reliable and useful results in ongoing experiments. 2. Do some research and describe two or three scenarios in which it would be preferable to use a stab tube vs. a growth plate. ( Hint : What do bacteria use to help them move? Can moIlity be used to help idenIfy many medically important pathogenic bacteria such as the Enterobacteriaceae family of bacteria?)

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“ Lab 2 Culturing & Aseptic Technique BIO250L ” 1. MoIlity: Stab tubes in clinical labs are used to disInguish between bacteria like E. coli and P. mirabilis based on their moIlity, crucial for diagnosing urinary tract infecIons. 2. PreservaIon: In research se_ngs, stab tubes are uIlized to preserve newly discovered bacteria with unique properIes, ensuring their long-term viability for future biotechnological studies.

BIO250 Lab 2 FINAL

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