...Laboratory Techniques: Pipetting Objectives: 1. To learn which pipettes are used for desired volumes. 2. To learn correct procedures for measurement and transfer of liquids. 3. To become familiar with the appearance of certain volumes in pipet tips. Introduction: Micropipettes are equipment used to transfer and measure very small amounts of liquid as low as 0.003 ounce (0.1 millilitres) in laboratory. Standard micropipettes used in laboratory settings feature a few basic parts including an adjustment dial and a plunger button. Normally, micropipettes are consisting several of components, such as volume adjustment dial and a digital volume indicator. These pipettes also get used to different types of disposable tips at the end of a shaft which is one part of special of micropipettes. Liquids are drawn into the disposable plastic tips fitted at the end of micropipette shafts instead of into the shafts themselves. There are 2 main buttons found on micropipettes, which are the plunger button and the tip ejector button. The pipette will immediately release the disposable tip at the end of the device by pushing the ejector button. Most of plungers of micropipette are designed to stop at two different positions. Pipette will draw the desired volume of liquid up into its disposable tip by depressing the plunger to the first stopping position and slowly releasing it. A micropipette's plunger is then pressing down further to the second stopping point...
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...Lab Report 1: Pipettes and Std Curve Ingrid Simons University of Texas at El Paso June 12, 2014 Introduction: Procedures that require very small volumes of DNA and reagents are performed with instruments such as, micropipettes and pipettes. These instruments are used when scientists need to accurately and precisely deliver small volumes of a liquid. Molecular biologists use very small volumes of liquid at times they are as small as 0.1 µL, in order to do these measurements the use micropipettes. The function of the micropipettes is to act as a plunger when the depressed by the thumb and by its release liquid is drawn into a disposable plastic tip. The liquid is released when the plunger is pressed again (The University of QueensLand). The tip is an important part of the micropipette, each tip is disposable and come in different sizes depending of the micropipette being used. There is three standard micropipettes, the P20 which measures 0.5-20μL, P200 that measures 20-200 μL, and P1000 that measures 100 μL to 1 mL. An important part of this laboratory was learning how to read the meniscus curve. A meniscus is a curve that is formed at the surface of a molecular substance, such as water, when it touches another material (USGS). Meniscus is caused by adhesion, a result of water’s high surface tension. The glass beaker attracts water molecules, and water molecules stay together, as a result many molecules gather to touch the glass. They will accumulate along the...
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...recording potential. Impedance of microelectrodes can play a very important role in the monitoring of low amplitude and high-resolution of the cells. Because of the smaller size of microelectrodes, it has high impedances in kilo ohms range. It is classified into two types a. Metal type b. Micropipette type a) Metal microelectrode: Metal microelectrodes are prepared by electrolytic etching the tip of fine tungsten to the desired size and dimension. Then the wire is coated almost to the tip with any type of insulating material. The metal-ion interface takes place where the metal tip contacts the electrolyte. The main features of metal microelectrodes are 1. It has excellent signal to noise ratio 2. It is strong enough and easy to penetrate 3. It has high biocompatibility b) Micropipette: The micropipette type of microelectrode is a glass micropipette with its tip drawn out to the desired size. The micropipette is filled with an electrolyte which should be compatible with the cellular fluids. A micropipette is a small and extremely fine pointed pipette used in making microinjections. A commercial type of micropipette is shown in Fig. 1.1.2. Figure1.1. 2: Micropipette type Electrodes Body Surface Electrodes: Body surface electrodes are placed in contact with the skin of the subject to obtain bioelectric potentials. Body surface electrodes are of many types and sizes. The various types of body surface electrodes are discussed below. Major body surface...
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...wipe the benches with Virkon to prevent microorganism growth. Disinfectants: low hazard. Wash hands after use. Ethanol: can cause a fire. Take care when using ethanol with the Bunsen. Bunsen: burns. Use carefully. Keep on yellow flame when not in use. E.coli: possibility of infections. Use a good antiseptic technique. Wear a lab coat. Cover cuts/ ware gloves. Depose of equipment in a bleach pot or bags for autoclaving. Hypothesis: I would expect to find that Dettol works better than Tesco’s disinfectant because it is said to work better than other leading brands. Equipment: Tweezers Borer Pipettes Suring (20Ym) Glass simmers Beaker of ethanol Bunsen burner E.coli 4 clean agar plates Virkon 30 Cm Ruler Micropipette Method: 1. 1 plate for each cleaner (2 plates used, other plates are in case a mistake is made) swamped the E.coli in the blue flame of the Bunsen burner. 2. Get a pipette of E.coli and up it on the plate. Flame the spreader (put it in ethanol then in the flame) be careful on to crack the spreader. Use the simmer to spread the bacteria around the agar plate 3. Flame borer (in the same way as the simmer but be careful if the borer is in the flame too long you can burn your hand. If you do run it...
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...Factors Affecting Enzyme Activity Analysis of Enzyme Activity: Catalase and Tyrosinase Introduction to Neurotransmitters: Acetylcholinesterase Abstract: A series of three labs were combined to observe the effects of some common biological enzymes: Catalase, Tyrosinase, and Acetylcholinesterase (AChE). Enzymes are catalytic proteins, that when present in a chemical reaction, are able to lower the action potential needed to create the reaction without being destroyed or altered themselves in the process. In Part A, my hypothesis stated that when Catalase is combined with H2O2 the rate of conversion to water and oxygen gas should double when 5-10 drops of enzyme is added and quadruple when 10-20 drops are added. In Part B, my hypothesis stated that increases in enzyme concentration or buffer pH the substrate of the final product will yield increased substrate, also, if the substrate concentration is increased then the enzyme will be less diluted, the buffer pH will increase, or there will be a temperature increase. In Part C, my hypothesis stated that tacrine will have an inhibitory effect on AChE, and that those effects will increase as the level of concentration increases. In all three labs I postulated that increases in temperature and concentration levels and would increase the rates and decrease time to form chemical reactions. We setup each lab with a series of increased concentrations and a control trial using DiH2O. We observed the results using the...
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...Using a p1000 micropipette, the gel was overlayed with a thin layer of DI water. Said steps were repeated for the second gel. The mixture was left at room temperature for 30 minutes allowing the mixture to completely polymerize. Any excess acrylamide in the mixing vessel was allowed to solidify before being thrown away. The top of the resolving gel was then rinsed with DI water and blot dried with a paper tower. Vinyl gloves were put back on for the next step. Using the same flask (cleaned) mentioned before, the following components of the stacking gel were mixed in said order: 3.4 ml deionized water, 0.83 ml 30% acrylamide mix, 0.63 ml 1.0 M Tris (pH 6.8), 0.05 ml 10% SDS, 0.05 ml 10% APS, 5 μl TEMED. The components were swirled and mixed using a plastic Pasteur...
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...12 ice cubes into the blender and then the liver (the cold temperature ensures that the enzyme does not denature while being blended) 3. Add approximately 200 ml of water into the blender using a graduated cylinder 4. Add 20 ml of sand into the blender (this will help smooth out the liver and make it easier for the liver to blend) 5. Blend all the materials for approximately 4 minutes and 50 seconds (you may need to blend the mixture for more or less time depending on the strength of your blender) Changing the temperatures: For 10⁰ C 1. Place a few ice cubes inside a 500 ml beaker and use a thermometer to achieve a temperature of 10⁰ C (cool using ice or warm using tap water) 2. Using a micropipette, transfer 4m cm3 of the catalase solution into a test tube 3. Using another micropipette, pour 3 cm3 of hydrogen peroxide into another test tube 4. Place both of the test tubes into the 500 ml beaker that was previously prepared and is at 10⁰ C 5. Using a thermometer, wait until both the solution and the mixture have reached 10⁰ C 6. Pour the catalase...
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...4/15/2015 BIO156 Lab 2 Print Lab 2 Biological Molecules and pH Introduction: Connecting Your Learning Biological organisms, like all things, are made up of elements. These elements combine to form organic molecules that create the basis for life. The main elements found in biological organisms include carbon (C), hydrogen (H), phosphorus (P), nitrogen (N), and oxygen (O). This lab describes how these elements form some of the most important molecules in life: carbohydrates, proteins, and fats. Resources and Assignments Multimedia Resources Required Assignments None Lesson 2 Lab 2 From the Lab Kit 7 test tubes Benedict's solution Biuret solution 15 micropipettes 10 pipettes Forceps pH test strips 4 unknown samples https://www.riolearn.org/content/bio/BIO156/BIO156_INTER_0000_v9/labs/lab02.shtml?print 1/21 4/15/2015 BIO156 Lab 2 Measuring spoons (teaspoon and tablespoon) 50 mL beaker Mortar and pestle Glass stirring rod 100 mL graduated cylinder Microscope slide Plastic funnel Test tube tongs Test tube rack 5 plastic cups Goggles Plastic gloves 1 tablespoon baking soda 1 tablespoon chicken soup 4 tablespoons sugar Required Materials 1 tablespoon cornstarch 2 tablespoons unflavored gelatin Student Provided Small saucepan Paper towel Oven glove or mitt Baking tray or aluminum foil (about an 18-inch sheet) Scissors Pencil Dime Microwave (optional) or Stove Permanent marker https://www.riolearn...
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...proteinase K followed by salting out and denaturing some proteins by SDS. * Recovery of DNA: * DNA is precipitated using ethanol or isopropanol in the presence of monovalent cations All DNA isolation protocols have a common goal in isolating high molecular weight DNA, which meet the following three criteria: 1) Purity should be high enough for further downstream applications such as restriction digestion. 2) DNA should be intact to give accurate and reproducible migration patterns on gel electrophoresis. 3) Yield should be high enough to obtain a required quantity of DNA from a reasonable amount of tissue. Materials and Methods: Materials Bench centrifuge, incubator, Falcon tubes, eppendorf tubes, micropipettes, micropipette tips gloves. DNA extraction Frozen blood samples were thawn 30ml of lysis buffer was added, mixed and kept on crushed ice for 15mins Centrifuged at 2000rpm for 10mins and supernatant decanted without disturbing the pellet Pellet was washed with 10ml SE buffer, vortexing briefly and...
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...salt used to create the samples was supplied by Carolina Biological Supply Company. The seven ranging salinities were prepared in 100 mL samples. Seven masses of salt were measured on a scale (0g, 3.5g, 5g, 7.5g, 10g, 12.5g, and 15g) and were corresponded to a beaker filled with 100 mL of distilled water. For each sample, four trials were replicated by filling 30 mL plastic vials with 25 mL of solution. 2400 cysts (one leveled scoop) were placed in each vial. After preparing four vials for each of the seven samples, the Brine Shrimp were given 24 hours to hatch. On the following day, each plastic vial was shook gently to mix the solution. 100 micrometers were extracted using a micropipette and were dispersed onto a slide. Each slide was examined under a compound light...
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...13000rpm. Many microcentrifuges only have settings for speed (rpm), not relative centrifugal force (rcf). Consequently, a formula for conversion is required to ensure that the appropriate setting is used in an experiment. The relationship between rpm and rcf is as follow, g=1.118 x 10ˆ-5 rs² where g is the relative centrifugal force, r is the radius of the rotor in centimeters and s is the speed of centrifuge in rpm. Objective: 1. To determine the relationship of speed of centrifugation with the coagulation of milk which separate from the mixture. 2. To separate coagulated milk from the mixture of fresh milk and acetic acid by centrifugation. Materials: Centrifuge, 15ml centrifuge tubes, 1.5ml microcentrifuge tubes, micropipettes, fresh milk, 50% of acetic acid, Falcon tube Procedure: 1. 5ml of fresh milk is drawn by the pipette into centrifuge tube which contains 50% of 10ml acetic acid. 2. Centrifuge tube was inverted gently for a few times to mix both solutions. 3. Milk sample was centrifuged...
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...Experimental Procedure Materials and Reagents The following materials and reagents were used throughout the experiment: P200 and P100 Pipetman micropipettes were used in this experiment. Fisher gel loading pipet tips were used. A Genesys 10S UV-VIS Spectrophotometer was used and had the ID number 2N3R294201. Two cuvettes (1.0cm path length) were used. A black Sharpie marker was used. A Fisher vortex mixer was used and had the ID number 060711069. A Fisher Scientific Isotemp was used and had the ID number 102NO562. Coomassie Brilliant Blue dye stain (2.5 g dye, 50% methanol, and 7.0% Acetic Acid in water) made on 4/22/2015 was used. Gel de-staining solution (50% Methanol/7.0% Acetic acid) made on 10/16/2013 was used. Two magnetic...
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...The baseline heart rate was obtained by setting the crayfish inside the container undisturbed with a piece of foil to give to simulate its natural environment. We recorded this for five minutes. The next reading was the stressed heart rate, for this we uncovered the crayfish and clicked start. To stressed the crayfish, we snapped our fingers and turned on the strobe light on our phones. This heart rate activity was recorded for five minutes. Serotonin was obtained from the refrigerator. Using the micropipette, we transferred one milliliter of solution into the container. Then we allowed the crayfish to sit with the foil cover for five minutes so the solution can equilibrate. After these five minutes, we clicked start and recorded crayfish’s response to serotonin. Clicked stop and marked the start and end points of the serotonin heart rate. The crayfish was removed from the container and the 100 ml of freshwater crustacean saline was...
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...Dye can be used on wet mounts but it’s completely optional. Some particular dyes can be used to discolour the DNA within the cell and lipids. Materials required Microscopic slides Coverslips Micropipettes Paramecium and pond water cultures Methods A clean and scratch free glass slide was taken and the slide was labelled with the name of the organism or the sample. In the middle of the slide 15-20 μl of culture was placed. Diagonally a clean coverslip was lowered and dropped from one side avoiding any air bubbles. The prepared slide was examined under the microscope firstly under 10× the following by 40× and 100× magnification. Observations were drawn and any mobility was recorded. After observations the glass slide and the cover slip were discarded into an appropriate...
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...Who Committed the Crime? #26 5 December 2013 Partner: C.C. Objective The reason for doing this experiment was to find out which suspect committed the crime of murder. The purpose and importance of this experiment was to take the blood samples found at the crime scene, analyze them and see whether the blood sample matched suspect A, the victim’s half sister or suspect B, the victim’s ex-boyfriend. Hypothesis Based on the information acquired, the educated guess of the suspect that committed the crime was Suspect A, the victim’s half sister. This was believed to be true because the victim and her half sister have never had a good relationship. Her jealously towards the victim was well known and they were seen in public one week prior to the incident arguing, which raises a red flag. Background The victim was a cheerleader in college and she was an honor student. She was murdered by multiple stabs and the guilty suspect disposed of the body in a dumpster near the library. Luckily, there was a trail of blood left at the scene and there was a blood sample that was connected to one of the suspects. The suspects narrowed down to her half sister who was very jealous of her and her ex-boyfriend. Similar to the procedure done in forensics the samples of the crime scene blood, the blood of suspect A and the blood of suspect B were combined with restriction enzymes and put through gel electrophoresis to determine which suspect’s blood matched the blood found at the...
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