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Explain why the following steps are essential during subculturing: a. Flaming the inoculating loop prior to and after each inoculation b. Cooling the inoculating loop prior to obtaining the inoculum c. Flaming the neck of the tubes immediately after unplugging and before replugging 2. Incubate the tubes, including control, in a bacteriological BOD incubator at 30C for 24 hours. Spread the bacteria over approximately a quarter of the plate, edge to edge. In subculturing the inoculating loop is used when you are using a liquid medium. Do not leave your loop in the incinerator for more than 10 seconds, you will destroy the loop! Inoculating agar plates, slopes and cultures. inoculate \ih-NAHK-yuh-layt\ inoculate. Figure 2.2. We've got the study and writing resources you need for your . Rub the inoculum onto a small area near the edge, sterilize the loop, and then go back and complete the streaking of the plate by using the technique illustrated in figure 9.1. Use the loop/needle to pick up organism from the broth. Place your loop in the mouth of the incinerator briefly for 2-4 seconds to sterilize it. Study Resources. Hold the cap with your little finger that is holding the loop/needle. Explain your answer. If using a sterile flat toothpick, hold the narrow end gently between your thumb and ring finger at a 10 to 20 angle to the medium, and use the wide end to streak the quadrants. We've got the study and writing resources you need for your . Grasp the tube cap with the little finger of your hand holding the inoculating loop and remove it from the tube. Answer (1 of 3): Inoculation loops need to be sterilised before and after every use to reduce the chances of contamination as much as possible to achieve the most accurate results possible within tests. Pick an isolated colony from the agar plate culture and spread it over the first quadrant (approximately 1/4 of the plate) using close parallel streaks or Insert your loop into the tube/culture bottle and remove some inoculum. Insert the inoculating loop into the culture (illustration 4, figure 2.1). using aseptic techniques . The loop must be cooled to prevent killing the bacteria. This leaves the little finger free to take hold of the screw cap/ cotton wool plug of the bottle/ test tube. tutor. bacterial . Before using either, the end of the wire must be sterilized by passing it slowly through the tip of the HarleyPrescott: Laboratory Exercises in Microbiology, Fifth Edition III. Alternately flame the mouth of both tubes and replace both caps to respective tubes. b. What is the purpose of subculturing? Do not let the loop touch any of the previously streaked areas. 4. Do not put down loop or wave it around! Carefully transfer a . Rotate the plate at 90 and remove the lid just like before just to fit to inoculating loop. Cool the inoculating loop prior to obtaining the bacterial sample 3. Subculture is used to prolong the life and/or expand the number of cells or microorganisms in the culture. First week only $4.99! 2. A loop is used when you are using a liquid medium. 4. 5. 1. Questions 1. - leaving the lid off too long, leaving the lid off test tube - unsterilized instruments, double dipping Click to see full answer. Thus, 200 l (0.2 ml) of water should be equal to 0.2 g. the st. lawrence seaway connects the great lakes, the . write. Flame the inoculating loop or needle (your preference) until it is red hot, allow to cool. How is it possible to contaminate a subculture? 6 Lift the lid a little of the Petri dish containing the inoculum with the left hand. Science Biology Q&A Library Explain why the following steps are essential during subculturing:a. Flaming the inoculating instrument prior to and after each inoculation.b. Which of these statements best describes the st. lawrence seaway? Inoculating loop (usually nichrome, a nickel-chromium alloy, or platinum; it may also be a single-use disposable plastic loop, which would be discarded between sectors rather than resterilized). When inoculating an agar slant, draw the loop containing the inoculum very lightly over the surface in a zigzag formation while being careful not to break the surface. Allow the loop to cool a few seconds to avoid killing the inoculum. What is the purpose of flaming the inoculating loop before and. Flame the mouth of the tube as shown in illustration 3, figure 2.1. S Hos is it possible to contaminate a subculture? In subculturing, when do you use inoculating loop and inoculating needle? Sterilize the inoculating loop in the bunsen burner by putting the loop into the flame until it is red hot. inoculationg loop is used to asceptically transfer microorganisms from broth, slant, or agar cultures to other media How is it possible to contaminate subculture? Touching a broth or a culture plate will. . Using a wider streak. In subculturing, when do you use the inoculating loop? the st. lawrence seaway provides an important trading route for the u. s, but has little value for canada. Alternately flame the mouth of both tubes. 5 Flame the loop and allow to cool. If using a loop or wooden stick, hold it like a pencil at the same angle. Also, why use an inoculating needle vs inoculating loop? In subculturing, when do you use the inoculating loop? Grab the inoculating loop far back on the handle as if you were going to write with it. Partially lift the lid of the plate culture and open it just enough to insert the inoculation loop. Why must you use aseptic les when carrying out subculturing? Use an analytical scale to measure water, making sure the minimum and maximum settings correspond to the intended volume. When you bring the loop out of the tube, be sure it holds some of the broth. Start your trial now! For example, use a P1000 to transfers 200 l of water to a weigh dish on the scale. Drag the loop lightly from the first section towards the second section and repeat the zig-zag pattern. First week only $4.99! Why was S. marcescens used in this exercise? Procedure . Culture Transfer Instru., Techniques, & Isolat. 2. Solution for Explain why the following steps are necessary during subculturing: Cooling the inoculating loop before touching the inoculum/culture ? The inoculating loop or needle is then streaked over an agar surface. Flame the loop and repeat step 8 in the last remaining section. Why was S. marcescens used in this exercise? 3. Cooling the inoculating instrument prior to obtaining the inoculum.d. 7. An inoculation loop is simple laboratory equipment mainly used by mycologists to transfer microorganisms to a growth media. Flame the opening of the broth before closing. There is some controversy as to the value of this action. 2. Allow the inoculating loop or needle to cool for 10 - 20 seconds. 6. Let it air dry for 15 min then cover with the mordant for 4 mins. Some examples of this are: - Leaving the lid of the culture off too long By not practicing aseptic technique (i.e. Using the same hand that is holding the inoculating loop, remove the caps from the two tubes, hold them between your fingers, and briefly flame the necks of the tubes over a Bunsen burner by passing them through the flame. If your lab uses sterile plastic loops, use a new loop for each transfer being mindful not to touch it on any surface. Therefore dust/particles in the air are less likely to fall into your tube. Holding the test tube caps in the hand as illustrated in Figure 1.1 on page 24.c. This action is called subculturing or passaging the cells. Hot air rises. Do not completely open the lid and expose the surface to the air. a. Sterilize the inoculating loop or needle by holding it in the flame of the gas burner, moving it through until the wire turns red. In biology, a subculture is a new cell or microbiological culture made by transferring some or all cells from a previous culture to fresh growth medium. The most isolated colony should be used when subculturing because any genetic mutations, such as antibiotic resistance, will have been gained by the most isolated colony (colonies). different environments and other possible variables. Regarding this, what is the purpose of flaming an inoculating loop? close. an inoculating needle; if a loop is present, it is an inoculating loop. learn. As demonstrated, use a sterilized inoculating loop to pick up one M. luteus colony (or a piece of a colony) and transfer it to the surface of the agar plate. 5. c : to introduce immunologically active material (such as an antibody or antigen) into especially in order to treat or prevent a disease. It also ensures that any liquid culture on the loop will run down into the flame. study resourcesexpand_more. The inoculating loop should be heated until it is hot enough to turn red, and then allowed to cool for a couple seconds. Pick up your sterile plate. Place the slide on top of a boiling water bath for 5 mins. . Fig 1: Inoculation of culture into agar slant 2. To process clinical specimens satisfactorily for bacteriological culture, consideration must be given to. learn. tutor. 4. In subculturing, when do you use the inoculating loop? The purpose of flaming an inoculating loop is to prevent or reduce the chances of cross-contamination of the cultures being inoculated. arrow_forward. minimice eanaminaticn 3. Remove the cap from the broth and flame the opening. 3. This SMI should be used in conjunction with other SMIs. Touch the loop to an area of the agar with no growth in order to cool down the loop. Start your trial now! Transfer a loopful of culture into the sterile broth with the sterile loop. a Carry out the transfer of cultures as quickly as possible, with tubes and plates open to the air for the minimum length of time.. b Normal practice is to open agar plates away from the body and without removing the lid completely from the base.. c In instances when the lid of the Petri dish may be removed for longer periods than normal, work very . For each portion of the streak (3 total per plate), flame-sterilize the inoculating loop just prior to use.Also, flame-sterilize the loop just after the final streak is performed in order to prevent contamination of the bench surface and as a consideration to others in the lab who may later use the inoculating loops. Using a wider streak. 7 Touch a single colony with the wire loop. Flame the opening of the broth before closing. 4 Hold the loop in the right hand. Explain why it is necessary to: a. Flame the inoculating loop before and after each inoculation. 4. Since water has a density of 1, then 1 ml of water is equivalent to 1 gram (g). A needle can be used instead of a loop to inoculate an agar slant by stabbing the needle containing the inoculum into the agar ( Fig 1). Inoculating loop (usually nichrome, a nickel-chromium alloy, or platinum; it may also be a single-use disposable plastic loop, which would be discarded between sectors rather than resterilized). Answer (1 of 2): Why reason for flaming the mouth of test tube? An inoculation loop is simple laboratory equipment mainly used by mycologists to transfer microorganisms to a growth media. 4. Transfer a loopful of culture into the sterile broth with the sterile loop. Let inoculating loop cool down for 5-10 seconds. Remove the cap from the broth and flame the opening. . "Flaming the mouth of a test tube creates an air flow. The purpose of flaming an inoculating loop is to prevent or reduce the chances of cross-contamination of the cultures being inoculated. Rinse throughly with water, add a paper towel on smear and soak paper with stain. On the initial region of the streak, many microorganisms are deposited resulting in confluent growth or the growth of culture over the entire surface of the streaked area. Hold both the culture and the broth tube in the left hand. Score: 4.5/5 (52 votes) . 2. Note: After you become proficient in streaking, you could visualize each petri dish divided into quarters instead of actually drawing lines. b. While using a wire loop, hold the handle of the wire loop close to the top, as you would hold a pen, at an angle that is almost vertical. Basic Laboratory and Culture Techniques 14. An inoculating loop is also known as a smear loop. If the loop is too hot, it will kill the bacteria and sizzle and melt the media that further promotes contamination. Cool the inoculating loop prior to obtaining the bacterial sample 3. b. write. Allow it to cool. Holding an inoculating loop between your thumb and index finger, insert the wire portion into the Bunsen burner flame, heating the entire length of the wire until it not flaming the mouth of the tube or inoculation tools). Using a wire loop. Sterilize your loop, by holding in the right hand, remove both plugs (or caps) from these tubes by grasping them between the fingers of right hand. How is it possible to contaminate a subculture? Add bacterium with loop to slide with 3 drops of distilled water, and spread it around. The inoculating loop is used when you are about to take a sample from a solid culture media (ex. Do not put down loop or wave it around. This is accomplished by using a volumetric inoculating loop calibrated to hold a specific volume of urine, preferably 0.001 mL or 0.01 mL. From your answer to question 3, can you say with a high degree of; Question: Questions 1. The loop is flamed once again before settling it down. Why must you use aseptic les when carrying out subculturing? Do not press so hard that the loop, stick or toothpick digs into the agar. Flame the inoculating loop or needle (your preference) until it is red hot, allow to cool. Answer (1 of 3): Inoculation loops need to be sterilised before and after every use to reduce the chances of contamination as much as possible to achieve the most accurate results possible within tests. Do not let the loop touch any of the previously streaked areas. The loop is used in the cultivation of microbes on plates by transferring inoculum for streaking. Using the other hand, flame the inoculating loop or needle over a Bunsen burner until the wire becomes red-hot. study resourcesexpand_more. Introduction . Solution for Explain why the following steps are necessary during subculturing: Cooling the inoculating loop before touching the inoculum/culture ? Procedure for inoculating a nutrient broth. Describe how to use the two most common types of pipettes. Score: 4.2/5 (23 votes) . On the other, write "mixed" to indicate that you're subculturing from the mixed culture broth to this plate. 6. Analytical Phase. Study Resources. Hold the cap with your little finger that is holding the loop/needle. 1. close. What is the purpose of flaming in the aseptic technique? This ensures that the heat kills the majority of lingering bacteria before. eSepiic . Also, it can be used to transfer microscopic organisms. The flaming of the loop at the points indicated is to effect . Get isolated colony of your yeast culture (if there is growth on 15-min and 30 min culture plate, do subculturing on each plate). Flame the inoculating loop before and after each inoculation. This quality guidance describes the methods of inoculating culture media and sub-culturing of organisms using aseptic techniques. 1 a : to introduce a microorganism into.b: to introduce (something, such as a microorganism) into a suitable situation for growth. A subculture can be contaminated by any substance error that introduces foreign matter into the culture. It is important to use a needle rather than an inoculating loop because the needle is used to transfer the specimen . The hot air will also creat. 5. Use the loop/needle to pick up organism from the broth. The inoculating loop is used to transfer culture from a broth. The flaming of the loop at the points indicated is to effect . Apply flame to sterilize the inoculating loop immediately after the transfer. 9 Replace lid of Petri dish. arrow_forward. the st. lawrence seaway provides an important trading route between the u. s. and mexico. Regarding this, what is the purpose of flaming an inoculating loop? The loop is sterilized by heating the loop in the blue flame of the Bunsen burner, between streaking . the st. lawrence seaway connects the st. lawrence river with the arctic ocean. Subculturing is also done to keep the microbial strains alive for scientific research. Inoculate the colony onto a new culture agar plate and do the streaking technique as shown in the video. Now use the loop to inoculate a fresh nutrient agar plate. 8 Withdraw loop. Pick up your sterile plate. 1,2: In subculturing, when do you use the inoculating loop?

in subculturing, when do you use the inoculating loop

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