procedure of streak plate method

The spread plate technique was used in lab #5 to obtain isolated colonies. A . Quadrant Streaking It is the most commonly used and the most preferred method where four equal-sized sections of the agar plate are streaked. The principle is to exhaust an initial deposit by making successive smears in different directions (hence the name of the technique: exhaustion by the quadrant method). Sterilize the loop in the flame. It is essentially a dilution technique that involves spreading a loopful of culture over the surface of an agar plate. Expand the streaks toward the center-third of the plate, back to the most recently streaked zone. a. 8. The object is to have the inoculation loop be slightly above the inner blue cone of the flame. It provides a simple and rapid method of diluting the sample by mechanical means. Serial Dilution Method. Streak Plate Procedure: Isolation of Bacterial Colonies Using the Quadrant Method The streak-plate procedure is designed to isolate pure cultures of bacteria, or colonies, from mixed populations . 3. 2. Turn the plate 90 degrees and drag the loop through the area you have just streaked two to three times and continue to drag the loop in a "zig-zag" formation in the remaining half of the plate without touching that area again. Repeat 4 and 5 step again untill the remaining area of the plate is filled with zig-zag formation. Microbiologists use the streak plate method every day to isolate colonies. Streaking is a technique used to isolate a pure strain from a single species of microorganism, often bacteria. This method is suitable for facultative, Microaerophilic, and anaerobic microorganisms. Through the streaking method, we can culture, isolate and study the individual colony of bacteria. 1. Microbiologists use the streak plate method every day to isolate colonies. Explain. Procedure: 1. at A in the diagram. The streak plate method is a rapid qualitative isolation method. There is a powerpoint posted that also has helpful informa:on on this procedure. When the loop is cool, it is dipped into an inoculum such as a broth or patient specimen containing many species of bacteria. Glass spreading rod b. Flame loop c. Agar plate d. This streak plate dilution method is achieved using the following steps: 1) Flame sterilize a wire loop. Using the streak plate method of isolation, obtain isolated colonies from a mixture of microorganisms. 2. Steps of Four-way Streak or quadrant streak: Sterilize the wire loop on flame and allow it to cool for a few seconds. Streak Plate Method Definition It refers to the pure culture technique that involves inoculating the separated colony from the mixed culture (either broth culture or directly from the culture plate/slant) over the solid agar surface by moving the sterilized inoculating loop back and forth. The streak plate method requires the number of organisms in the inoculums be reduced. Rotate the plate about 60 degrees and spread the bacteria from the end of the first streak into a second area using the same motion in step 6. The streak plate method is used as an isolation technique, which ensures that after 4-5 streaks you have isolated colonies. Turn the plate 90 degrees and drag the loop through the area you have just streaked two to three times and continue to drag the loop in a "zig-zag" formation in the remaining half of the plate without touching that area again. Turn the plate 90 degrees and streak across the original line to evenly disperse the bacteria across the agar. 7. The inoculation of the culture is made on the agar surface by back and forth streaking with the inoculation loop over the solid agar surface. Turn the plate 90 degrees and drag the loop through the area you have just streaked two to three times and continue to drag the loop in a "zig-zag" formation in the remaining half of the plate without touching that area again. A sample is taken from a colony and a microbiological culture is grown on the new plate in order for the organism to be identified properly. Due to a limited population of bacteria in the streaking method, pure culture isolation is quite easier than the pour plate and the spread plate method. 1) Create liquid overnight culture in BHI with single colony of L. monocytogenes. Sample preparation Quadrant Streak Tips Use the entire surface of the plate, not just the middle of the plate. Significance of Streak Plate Method It was originally developed by the two bacteriologists, Loeffler and Gaffkey in the lab of Robert Koch (Father of Bacteriology). The dilution or isolation by streaking method was first developed by Loeffler and Gaffky in Koch's laboratory, which involves the . Answer (1 of 2): This method is used to dilute a concentration of microorganisms, making them easier to work in lower concentrations. What is the max dilution needed to make certain that a countable plate (25-250 colonies) is achieved for a sample of pasteurized milk for which the max expected count is 100,000 CFU/ml? Follow these best practices to get your streaking technique down to a science. The streak-plate procedure is designed to isolate pure cultures of bacteria, or colonies, from mixed populations by simple mechanical separation. Turn the plate 90 degrees. Go To Results of Streak Plate Lab Procedures POUR PLATE CULTURE METHOD FOR THE ISOLATION OF MICROORGANISM IN LABORATORY REQUIREMENTS FOR THE POUR PLATE TECHNIQUE 24 hours old nutrient broth culture of two or more bacteria (Mixed Culture) or Sample/Specimen. 1. Limitations of Pour plate method. Streak Plate Procedure: Isolation of Bacterial Colonies Using the Quadrant Method The streak-plate procedure is designed to isolate pure cultures of bacteria, or colonies, from mixed populations by simple mechanical separation. Remove only one colony or a barely visible amount of cells from a plate or slant. 16.13). Repeat the procedure. Single colonies are comprised of millions of cells growing in a cluster on or within an agar plate ( Figure 1 ). Loss of viability of heat-sensitive organisms coming into contact with hot agar. 2. Samples can then be taken from the resulting colonies and a culture can be grown on a new plate so that the organism can be identified, studied, or tested. With the aid of a sterile nichrome wire loop, remove a sample of bacterial culture from the culture tube. 1 Begin by connecting the Bunsen burner to a fuel source and igniting it. Streak Plate Method of Isolation The most common way of separating bacterial cells on the agar surface to obtain isolated colonies is the streak plate method. It provides a simple and rapid method of diluting the sample by mechanical means. Sterilize the loop again in the flame and cool it at the edge of the agar away from the bacteria in the plate that you just streaked. The streak plate method will require constant use of the streaking loop. Those 28 colonies were subcultured . About Press Copyright Contact us Creators Advertise Developers Terms Privacy Policy & Safety How YouTube works Test new features Press Copyright Contact us Creators . 1. flame loop and let cool 2. lift lid of petri plate 3. streak across streak a and into quadrant 2 4. streak within quadrant 2 5. close petri plate lid 6. rotate plate and flame loop A student streaks a plate using inoculum from a broth culture tube. Hold it there for several seconds. Reflame the loop and cool it. Procedure Sterilize the inoculating loop in the bunsen burner by putting the loop into the flame until it is red hot. 6. For accurate counts, the optimum count should be within 30-300 colonies/plate. Label the plate and incubate it inverted. Thus, one must be careful to score these . A small amount of mixed culture is placed on the tip of an inoculation loop/needle and is streaked across the surface of the agar medium (Fig. Show the inoculating loop to the flame and keep it. Streak Plate. Streak plates 1.With this method, a small amount of sample is placed on the side of the agar plate (either with a swab, or as a drop from an inoculating loop if the sample is a liquid). Embedded colonies are much smaller than those which happen to be on the surface. The streak plate is a qualitative isolation method; quadrant streaking is mostly done to obtain pure colonies. Isolated colonies are clones that arise from a single cell, which ensur. Streak plate method: Principle, Purpose, Procedure and results Overview: Streak plate technique is In order to be able to properly identify a bacterial species, it is necessary to obtain well isolated colonies. 3) Perform serial dilutions down to 10^-10 and plate 10uL (or 100uL I forget) of each one spreading with a loop on . View Streak plate method Principle_ Purpose from APPLIED SC as114 at MARA University of Technology. Colonies develop sub-surface. The word "streak" means "a long, thin line." In the streak plate method procedure, a sample is spread in a petri dish in the shape of a long, thin line over the surface of solid media. Stretch the streaks into the third quarter of the plate and return to the last streaked location. Streak Plate Method- Principle, Methods, Significance, Limitations Streak Plate Method- Principle, Methods, Significance, Limitations In microbiology, streaking is a technique used to isolate a pure strain from a single species of microorganism, often bacteria. Loeffler and Gaffkey introduced the streaking method. As you might guess, the purpose of streaking for isolation is to produce isolated colonies of an organism on an agar plate. why is streak plate preferred over spot inoculation in Blood Agar procedure? (2, 5, 7 . Using the streak plate method of isolation, obtain isolated colonies from a mixture of . Sterilize the loop in the flame. Quadrant Streak Plate Method Step 1: Obtain the proper media plate for your bacterial sample, such as an LB agar plate; label the bottom with your name, date and bacterial species, and set up your materials in a sterile environment such as a laminar flow hood or next to a Bunsen burner with a clean bench top. Carefully transfer a loopful to the agar plate by making a single streak down the plate. Use the simulated agar surface below to practice the streak pattern using a pen or pencil. As the loop is streaked across the agar surface, more and more bacteria are rubbed off until individual separated . Nutrient Agar Medium Six 9 ml Sterile Water Blanks Sterile Petri plates Marker Graduated pipette (1ml) PROCEDURE OF POUR PLATE TECHNIQUE Which of the following is used in the spread plate and not the streak plate method? It requires strict maintenance. The purpose of the streak plate is to sample and isolate/dilute the bacteria into individual colonies What is the purpose of the streak plate? Watch the videos and go over the websites posted in this folder. A loopful of inoculum is taken and streaked such that the first quadrant contains the highest concentration of the inoculum, followed by the second quadrant, third quadrant, and fourth . Incubate for 18-24 hours and check at regular intervals. Procedure. The inoculation loop is first sterilized by passing it through a flame. PROCEDURE 1. 2. It is also known as a four-quadrant streak. 4. Streak Plate Method: This method is used most commonly to isolate pure cultures of bacteria. Dip the loop in the Liquid sample containing a mixture of bacteria. At this point, the loop is sterile and extremely hot. The procedure includes a dilution technique which requires spreading a loopful of culture over the agar plate surface. Streak Plate Method of Isolation. The most common way of isolation of pure culture techniques cells on the agar surface to obtain isolated colonies is the streak plate. The inoculum is streaked over the. The pour plate method of counting bacteria is more precise than the streak plate method.On average, it will give a lower count as heat-sensitive microorganisms may die when they come in contact with a hot molten agar medium. The spread plate culture method is one of the commonly used culture technique for the isolation of microorganisms, especially the bacteria, in the laboratory. 5. Pick off isolated colonies of microorganisms growing on a streak plate and aseptically transfer them to sterile media to obtain pure cultures. Sterilize the loop again using the procedure in step 7. In a streak plate, dilution is achieved by first spreading the specimen I have done 2 different method to isolate mercury resistant bacteria from Soil. 2.A sterile loop (flamed until red hot, then cooled by touching the agar away from the inoculated sample) is then used to spread the bacteria out in . One of the most important techniques you will learn this semester is how to streak for isolation. Objectives of Streak Plate Method Principle of Streak Plate Method Types Show the loop to flame and then let it cool down.Then go to the edge of each line of the first quadrant and extend this to second quadrants sets.1 Again, sterilize the loop and in the same manner streak the third set of quadrants. Streaking may seem like Microbiology 101, but bad habits can lead to errors and contamination. Turn the plate 90 degrees. Fire up the loop again and let it cool. Allow it to cool. 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. 3 Remove the loop from the flame. Label first, streak second. T-Streak Method 1. Streak Plate Procedure: Isolation of Bacterial Colonies Using the Quadrant Method. For more information, visit http://www.bio-rad.com/yt/4/biotech-lab-textbook.This video demonstrates the streak plate method for isolating a single colony of. The streaking process will dilutes out the sample that was . 3. 3. more of the agar. This video explains about the different streak plate techniques and the purpose of the technique. 1. A wrong streaking method can ruin your plate. This is to make sure that the individual cells fall apart on the agar medium surface so as separation of the different species takes place. Streak Plate Method and isolation techniques . This technique is much known for the isolation and culturing of bacteria. Streaking is a technique for the isolation into a pure culture of the organisms (mostly bacteria), from a mixed population. 5. Starting at A, you heat your loop to a glow and let it cool, then inoculate your culture and spread the culture in back and forth streaks down about a fifth the wa. Repeat the procedure. This is a simple & rapid method. streak-plate method on agar. Table of Contents What is Streak Plate Method? It is also referred to as the "four-quadrant . To make sure that it is sterile and to make sure that only the cells your loop is touching are smeared into the next quadrant You don't need a huge chunk. Single colonies are comprised of millions of cells growing in a cluster on or within an agar plate (Figure 1). 1. . Procedure of Spread Plate Method The general procedure of the spread plate method can be summarized as: Arrange all the requirements, put on the PPE, sterilize the work surface, and allow all the samples and media to come to room temperature if were refrigerated. Eventually, it will wear out and you need to change it every once in a while. Answer (1 of 3): The streak plate method and spread plate method are used with different objectives. 3) Dip the loop into the sample. Quadrant streak is better for colony isolation as you start out with a much denser same and then isolate it down, . Turn the plate 90 and lightly sweep the loop 1-2 times through the inoculated area, then streak into the next quadrant without overlapping the previous streaks. Streaking may seem like Microbiology 101, but bad habits can lead to errors . The pour Plate Method technique was established in the laboratory of Robert Koch and is still being used widely since his period. Arrange the next steps of the procedure in their correct order. a method of handling microbial cultures, patient specimens, and other sources of microbes in a way that reduces contamination and prevents infection of the handler or others who may be exposed colony visible mass of bacteria growing together on an agar plate or slant. OCD = CFU/loop volume OCD = 150 CFU/0.001 mL Clearly labeling your plates to identify the microorganism only takes a [] This allows for individual colonies to be isolated from other colonies. All of these methods dilute or "thin out" a heavy population of bacteria across an agar surface. 1. Figure 2: Streak Plate Method Procedure. Bacteria are obtained from a mixed culture with a. sterile loop or wire and diluted within three sectors on a petri dish containing appropriate growth medium. Oxidase test is an important differential procedure that should be performed on all gram . Streaking is a method that isolates a pure strain from a species of bacteria. colonies are derived from a single microorganism colony forming units (CFUs) Principle The streak plate technique is essentially a method to dilute the number of organisms, decreasing the density. It is simple, less resource-consuming, easy, and economical; however, it requires. The techniques commonly used for isolation of discrete colonies initially require that the number of organisms in the inoculums be reduced. Record your counts in CFU/mL. Streak only one loopful of a broth culture. 2 Insert the loop into the flame at a vertical angle. 2) Cool the loop by touching the auger plate. Isolating bacteria: quadrant streak plate method lab Part 1: Terminology and procedure questions from videos 1. The general procedure of the streak plate method can be summarized as: Arrange all the requirements, put on the PPE, sterilize the work surface, and allow all the samples and media to come to room temperature if were refrigerated. Preparation for pour plate method is time consuming compared with streak plate/and or spread plate technique. Reflame the loop and cool it. 2. In Streak plate technique, a sterilized . Streak Plate Method of Isolation Purpose The streak plate technique is the most widely used method of obtaining isolated colonies from a mix of cultures. You will be answering ques:ons on this video, so please go over the ques:ons first. To ensure a countable plate, plate a series of dilutions. After incubation, as a result of this dilution technique, The nichrome wire loop should be sterilised using the Bunsen burner flame. 3 2 4 1 Inoculation of a streak plate 1. In this technique, a serially diluted specimen containing 2 or more bacteria or microbe (Mixed culture) is used which is spread over the solidified agar media plates as a thin layer with . A Loopful of culture sample is streaked on the agar surface in area 1 Now again wire loop is flame sterilized and now the Petri plate is rotated at 90 degrees. Flame the loop. Reflame the loop and cool it. This is accomplished in stages, as will be demonstrated in lab before you try it yourself. : Quadrant Streaking, T-Streaking, Continuous Streaking, and Radiant Streaking. The streak-plate method is an example of a. solid dilution technique. (area 3), extend the streaks into the center fourth of the plate (area 4). General Procedure There is a label on the bottom of the Petri dish. 2. Once again, flame your loop. RESULTS 1. Streak Plating. Types of Streak Plate Method. The streak plate method can be time-consuming, especially if you are going to prepare a large sample size. 2. Streak plate method: From my mix culture in Nutrient Broth, I got 28 colonies. 1. The two most common streak patterns are the three sector T streak and the four sector quadrant streak. . If the sample is very concentrated then dilution can be helpful to get the isolated colonies. 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. The inoculum for a streak plate could come from any type of source, for example clinical specimen, sedimented urine, environmental swab, broth, or solid culture. Quadrant Streaking: This is the most common method of streaking, where the petri dish is divided into four quadrants and then inoculated. 4. Repeat the technique for the fourth set of quadrants. Streak plate technique is one of the isolation techniques u. The Streak Plate technique for the isolation of microorganism is the most practical method of obtaining discrete and well-developed colonies of the microbe in pure cultures. The goal of streak plate method is to dilute the cells by spreading them out over the surface of the agar. Streak Plate Method - Explained What is Streak Plate Method ? Typical labels include the organism's name and the date. Types of Streak Plate Method Based on the pattern of streaking, the streak plate method can be classified into 4 types, viz. Streak plate - the original culture is directly diluted across an agar surface using and inoculating loop. . This is useful when you need to separate organisms in a mixed culture or when you need to study the colony morphology of an organism.

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