Biotechnology Explorer - Bio-Rad
araCoripGLOGFPblaBiotechnologyExplorer pGLO BacterialTransformation KitCatalog #166-0003EDUexplorer.bio-rad.comSee individual components for storage temperature.Duplication of any part of this document is permitted for classroom use only.Please visit explorer.bio-rad.com to access our selection of language translationsfor Biotechnology Explorer kit curricula.For technical support call your local Bio-Rad office, or in the U.S., call 1-800-424-6723
How can jellyfish shed light on the subject?One of the biggest challenges for first-time students of biotechnology or molecular biologyis that many of the events and processes they are studying are invisible. TheBiotechnology Explorer program has a solution: a gene from a bioluminescent jellyfish and itsGreen Fluorescent Protein—GFP. GFP fluoresces a brilliant green when viewed with ahand-held long-wave ultraviolet light (such as a pocket geology lamp).The gene for GFP was originally isolated from the jellyfish, Aequorea victoria. The wild-typejellyfish gene has been modified by Maxygen Inc., a biotechnology company in SantaClara, California. Specific mutations were introduced into the DNA sequence, which greatlyenhance fluorescence of the protein. This modified form of the GFP gene has been insertedinto Bio-Rad’s pGLO plasmid and is now available exclusively from Bio-Rad for educationalapplications.GFP is incredibly bright. Using pGLO to transform bacteria, students can actuallyobserve gene expression in real time. Following the transformation with Bio-Rad’s GFPpurification kit, students purify the genetically engineered GFP from their transformed bacteriausing a simple chromatography procedure. The entire process is visible using the hand-heldUV lamp.Guided InvestigationThe intent of this curriculum is to guide students through the thought process involved in alaboratory-based scientific procedure. The focus here is not so much on the answer orresult, but rather on how the result was obtained and how it can be substantiated by carefulobservation and analysis of data. This is referred to as a guided inquiry-based laboratoryinvestigation.At each step along the way, student understanding of the process and the analysis of datais stressed. Instead of providing students with explanations or interpretations, the StudentManual poses a series of questions to focus and stimulate thinking about all aspects of theinvestigation. Answers are provided in the Instructor’s Answer Guide.Student involvement in this process will result in an increased understanding of thescientific process and the value of proceeding into a task in an organized and logicalfashion. Furthermore, we are expecting that students who engage in this type of processwill start to develop a more positive sense of their ability to understand the scientificmethod.Bio-Rad’s GFP-based curriculum is unique and has generated an unprecedented level ofexcitement among science educators. We strive to continually improve our curriculum andproducts. Your input is extremely important to us. We welcome your stories, comments,and suggestions.Biotechnology Explorer TeamBio-Rad Laboratories6000 James Watson DriveHercules, CA 94547Biotechnology Explorer@bio-rad.com
Create context. Reinforce learning. Stay current.New scientific discoveries and technologiescreate more content for you to teach,but not more time. BiotechnologyExplorer kits help you teach moreeffectively by integrating multiplecore content subjects into asingle lab. Connect conceptswith techniques and putthem into context withreal-world scenarios. Genetic engineering oforganisms Use of experimental controls Interpretation of experimentalresults Calculate transformationefficiencyEnvironmentaland HealthScience Genetically modifiedorganisms (GMOs) GMOs in research, medicine,nutrition, and bioremediation Global challenges of GMOs mationChemistryof LifeKit Prokaryotic cell structureand cell division Selective growth media Bacterial metabolism Antibiotic selection andresistance genes Selection mechanisms Adaptation to environment Bacterial conjugation andgene transmissionCell andMolecularBiologyEvolution DNA structure, function,and chemistry Chemical properties ofbiological molecules Effects of temperature andpH on biochemical reactionsGenetics DNA RNA protein traitBacterial transformationThe lac operonCreating genetically engineeredorganisms (GMOs) Structure and function of genes Gene regulation and transcriptionfactors
Table of ContentsInstructor’s GuidePageIntroduction to Transformation .1The pGLO System .1Kit Inventory Checklist.2Implementation Timeline .5Safety Issues .5Lesson Points to Highlight.6General Laboratory Skills .6Experimental Points – Optimizing your pGLO Lab Experiment .7Conceptual Points .8Instructor’s Advance Preparation Overview.11Workstation Checklist .11Instructor’s Advance Preparation Guide .13Quick Guide (Graphic Laboratory Protocol).18Instructor’s Answer Guide .20Student ManualLesson 1Introduction to Transformation.32Focus Questions .33Lesson 2Transformation Laboratory .36Review Questions .42Lesson 3Data Collection and Analysis.43Analysis of Results .44Review Questions .45Lesson 4Extension Activity: Calculate Transformation Efficiency .47AppendicesAppendix AHistorical Links to Biotechnology .53Appendix BGlossary of Terms.57Appendix CBasic Molecular Biology Concepts and Terminology .59Appendix DGene Regulation .64Appendix EPhotodocumentation of pGLO Plates using Vernier’s BlueViewTransilluminator.66Appendix FReferences .67
Introduction to TransformationIn this lab, your students will perform a procedure known as genetic transformation.Genetic transformation occurs when a cell takes up (takes inside) and expresses a newpiece of genetic material—DNA. This new genetic information often provides the organismwith a new trait which is identifiable after transformation. Genetic transformation literallymeans change caused by genes and involves the insertion of one or more gene(s) into anorganism in order to change the organism’s traits.Genetic transformation is used in many areas of biotechnology. In agriculture, genes codingfor traits such as frost, pest, or drought resistance can be genetically transformed intoplants. In bioremediation, bacteria can be genetically transformed with genes enablingthem to digest oil spills. In medicine, diseases caused by defective genes are beginning tobe treated by gene therapy; that is, by genetically transforming a sick person’s cells withhealthy copies of the defective gene that causes their disease.Genes can be cut out of human, animal, or plant DNA and placed inside bacteria. For example,a healthy human gene for the hormone insulin can be put into bacteria. Under the right conditions,these bacteria can make authentic human insulin. This insulin can then be used to treatpatients with the genetic disease, diabetes, because their insulin genes do not function normally.The pGLO SystemWith the pGLO transformation kit, students use a simple procedure to transform bacteriawith a gene that codes for Green Fluorescent Protein (GFP). The real-life source of thisgene is the bioluminescent jellyfish Aequorea victoria, and GFP causes the jellyfish tofluoresce and glow in the dark. Following the transformation procedure, the bacteriaexpress their newly acquired jellyfish gene and produce the fluorescent protein which causesthem to glow a brilliant green color under ultraviolet light.In this activity, students will learn about the process of moving genes from one organismto another with the aid of a plasmid. In addition to one large chromosome, bacteria naturallycontain one or more small circular pieces of DNA called plasmids. Plasmid DNA usuallycontains genes for one or more traits that may be beneficial to bacterial survival. In nature,bacteria can transfer plasmids back and forth, allowing them to share these beneficialgenes. This natural mechanism allows bacteria to adapt to new environments. The recentoccurrence of bacterial resistance to antibiotics is due to the transmission of plasmids.Bio-Rad’s unique pGLO plasmid contains the gene for GFP and a gene for resistance to theantibiotic ampicillin. pGLO also incorporates a special gene regulation system that can be usedto control expression of the fluorescent protein in transformed cells. The gene for GFP can beswitched on in transformed cells simply by adding the sugar arabinose to the cell’s nutrientmedium. Selection for cells that have been transformed with pGLO DNA is accomplished bygrowth on antibiotic plates. Transformed cells will appear white (wild-type phenotype) on platesnot containing arabinose, and fluorescent green when arabinose is included in the nutrient agar.The unique construction of pGLO allows educators and students, for the very first time, to easilyexplore mechanisms of gene regulation (Appendix D) and genetic selection. And, the entireprocess is observable with an inexpensive long-wave UV lamp or with the provided pen-light.In order for your students to gain the most from this experiment, they should know whata gene is and understand the relationship between genes and proteins. For a moredetailed discussion of these and other basic molecular biology concepts and terms, refer tothe review provided in Appendix B.This pGLO transformation kit provides the opportunity for additional activities involvingpurification of the recombinant fluorescent protein from transformed bacteria using the GFPchromatography kit (catalog # 166-0005EDU) and the separation of proteins expressed inE. coli, such as the GFP protein by using the pGLO SDS-PAGE Extension kit (catalog#166-0013EDU.)1
Kit Inventory Check ( ) ListThis section lists the components provided in the bacterial transformation kit. It alsolists required accessories. Each kit contains sufficient materials to outfit 8 studentworkstations. Use this as a checklist to inventory your supplies before beginning theexperiments. All kit components can be stored at room temperature until use.Kit ComponentsE. coli HB101 K–12, lyophilizedPlasmid (pGLO), lyophilized, 20 µgAmpicillin, lyophilized, 30 mgL ( ) Arabinose, lyophilized, 600 mgTransformation solution (50 mM CaCl2, pH 6.1), sterile, 15 mlLB nutrient broth, sterile, 10 mlLB nutrient agar powder, sterile (to make 500 ml), 20 gPipets, sterile, individually wrappedInoculation loops, sterile, 10 µl, packs of 10 loopsPetri dishes, 60 mm, sterile packs of 20Multicolor microcentrifuge tubes, 2.0 mlFoam micro test tube holdersUV pen lightInstruction manual (Available online orprinted manual available by request)Required Accessories – Not included in this kitClock or watch to time 50 secMicrowave ovenTemperature controlled water bath, 1–6 liter(catalog # 166-0504EDU)*Thermometer that reads 42oC1 L flask500 ml graduated cylinderDistilled water, 500 mlCrushed ice, not cubed ice, and containers (foam cups work well)10 ml of bleach (household variety), 10% solutionPermanent marker pensNumber/Kit( )1 vial1 vial1 vial1 vial1 bottle1 bottle1 pouch508 pks2 pks6081 1 Number/Kit( )11 111111–810 ml4–8 * If a temperature controlled water bath is not available, obtain a container (foam is best) for hot waterand use a hot plate or hot tap water to get the water to 42 C.Optional AccessoriesNumber/Kit( )VortexerMicropipets, adjustable volume, 2–20 µl(catalog #166-0506EDU or 166-0551EDU)1 1 Parafilm laboratory sealing film2–20 µl pipet tips11 37 C incubator oven (catalog #166-0501EDU)**1 Vernier Blue Digital BioImaging System (BL-DBS)1 ** If an incubator oven is not available, try using an electric blanket or construct a homemadeincubator with a cardboard box and a low voltage light bulb inside. Otherwise incubate agar plates48 hours to 72 hours at ambient room temperature (see General Lab Skills–Incubation).2
Catalog #Product Description166-0555EDUpGLO Bacterial Transformation Kit Refill Package166-0405EDUpGLO Plasmid, 20 µg, lyophilized166-0406EDUArabinose, 600 mg, lyophilized166-0407EDUAmpicillin, 30 mg, lyophilized166-0408EDUE. coli strain HB101 K-12, lyophilized166-0409EDUTransformation Solution, 15 ml166-0421EDULB broth, 10 ml166-0600EDULB Nutrient Agar Powder, 20 g, makes forty, 60 mm agar plates166-0472EDULB Nutrient Agar Powder, 500 g, makes one thousand,60 mm agar plates166-0479EDUJellyfish Foam Floating Racks, 8 racks with 12 microcentrifugetube wells166-0500EDULong-Wave UV Lamp, 1166-0530EDUUV Pen Light, 1166-0470EDUPetri Dishes, 60 mm sterile, 500166-0471EDUInoculation Loops, sterile, 80166-0474EDUDisposable Plastic Transfer Pipets, sterile, 500166-0480EDUDisposable Plastic Transfer Pipets, nonsterile, 500166-0473EDUColored 1.5 ml Micro Test Tubes, 6 colors, 600223-9480EDUEZ Micro Test Tubes, 1.5 ml, natural, 500223-9430EDUEZ Micro Test Tubes, 2.0 ml, natural, 500166-0033EDUInstruction Manual (Available online or printed manual availableby request)3
National Science Standards and pGLO Transformation.Science as Inquiry Design and conduct scientific inquiry.Students will understand the need for controls for the experiment. Use technology and mathematics to improve investigations and communications.Students use mathematics to calculate transformation efficiency. They can evaluatedifferences in techniques based on efficiencies.Life Science StandardsThe Cell Cells have particular structures that underlie their functionsStudents should understand the role and function of the cell membrane and howprocedures in the lab are designed to get the plasmid across the cell membrane. Most cell functions involve chemical reactions.Transformation solution contains calcium chloride. Students should understandhow this salt dissociates in solution to surround the charged DNA molecules. Cells store and use information to guide their functions.Students provide new information to the cell that allows it to make new proteins. Cell functions are regulated.The production of the GFP protein is regulated by the sugar arabinose In all organisms, the instructions for specifying the characteristics of the organismare carried in DNA.The plasmid introduced to the cell is a circular, autonomously replicating piece ofDNA. Changes in DNA (mutations) occur spontaneously at low rates.Bacteria have the ability to change and survive in the presence of antibiotics.Biological Evolution Species evolve over time.Bacteria evolved with plasmids to provide new genes and new proteins for survival.Interdependence of Organisms Living organisms have the capacity to produce populations of infinite size, butresources are finite.Bacterial growth is limited as food depletes and waste increases in the petri dish.Science and Technology Understand about science and technologyRecombinant DNA Technology allows us to put genes from one species intoanother and have that species produce a new protein.4
Implementation TimelineEach of the three lab sessions is designed to be carried out in consecutive 50 min periods.The detailed lab protocol can be found in the Student Manual.Suggested laboratory schedule for the studentsDay 1Setting the StageLecture and discussionStudent considerations 1–4Day 2Transformation LaboratoryTransform cells and spread platesStudent laboratory focus questionsDay 3Data Collection and AnalysisObserve transformants and controlsAnalyze and interpret resultsStudent considerationsDay 4Extension ActivitiesCalculate transformation efficiencyGFP chromatography kit(catalog #166-0005EDU)pGLO SDS-PAGE Extension kit(catalog #166-0013EDU)Safety IssuesSome countries outside the U.S. may require a special license to use this kit. Please refer toyour country’s legislative authorities for proper guidelines.The Escherichia coli bacteria HB101 K-12 strain contained in this kit is not a pathogenicorganism like the E. coli strain O157 H7 that has sometimes been implicated in food poisoning.HB101 K-12 has been genetically modified to prevent its growth unless grown on anenriched medium. However, handling of the E. coli K-12 strain requires the use of standardMicrobiological Practices. These practices include, but are not limited to, the following.Work surfaces are decontaminated once a day and after any spill of viable material. Allcontaminated liquid or solid wastes are decontaminated before disposal. All persons mustwash their hands: (i) after they handle material containing bacteria, and (ii) before exitingthe laboratory. All procedures are performed carefully to minimize the creation of aerosols.Mechanical pipeting devices are used, mouth pipetting is prohibited; eating, drinking,smoking, and applying cosmetics are not permitted in the work area; wearing protectiveeyewear and gloves is strongly recommended.If an autoclave is not available, all solutions and components (loops and pipets) that havecome in contact with bacteria can be placed in a fresh 10% bleach solution for at least20 min for sterilization. A shallow pan of this solution should be placed at every lab station.No matter what you choose, all used loops and pipets should be collected for sterilization.Sterilize petri dishes by covering the agar with 10% bleach solution. Let the plate stand for1 hr or more, and then pour excess plate liquid down the drain. Once sterilized, the agarplates can be double bagged and treated as normal trash. Safety glasses are recommendedwhen using bleach solutions.Ampicillin may cause allergic reactions or irritation to the eyes, respiratory system, andskin. In case of contact with eyes, rinse immediately with plenty of water and seek medicaladvice. Wear suitable protective clothing. Ampicillin is a member of the penicillin family ofantibiotics. Those with allergies to penicillin or to any other member of the penicillin familyof antibiotics should avoid contact with ampicillin.5
Please obtain the Material Safety Data Sheets (MSDS) available from Bio-Rad by calling800-424-6723 in the United States, or see www.bio-rad.com for further information onreagents in this kit. Please consult your local environmental health and safety regulationsfor proper disposal.Ultraviolet (UV) LampsUltraviolet radiation can cause damage to eyes and skin. Shortwave UV light is more damaging than long-wave UV light. The Bio-Rad UV lamp recommended for this module is longwave. If possible, use UV-rated safety glasses or goggles.Lesson Points to HighlightThis section describes experimental and conceptual points which may prove challengingto students. These points are extremely important to the overall outcome of the activity.Instructors should alert their students' attention to these points, and when possible,demonstrate the technique before the students attempt the procedure.The most important thing for students to do is to put the correct components in the correcttubes and onto the correct plates. So, marking the tubes clearly and being prepared andorganized is crucial for a smooth execution of the experiment. The Quick Guide is providedto organize the activity. This graphic laboratory protocol provides visual depictions ofall laboratory steps used in the transformation procedure.General Laboratory SkillsSterile TechniqueWith any type of microbiology technique (such as working with and culturing bacteria), it isimportant not to introduce contaminating bacteria into the experiment. Because contaminatingbacteria are ubiquitous and are found on fingertips, benchtops, etc., it is important to avoidthese contaminating surfaces. When students are working with the inoculation loops, pipets,and agar plates, you should stress that the round circle at the end of the loop, the tip of thepipet, and the surface of the agar plate should not be touched or placed onto contaminatingsurfaces. While some contamination will likely not ruin the experiment, students would benefitfrom an introduction to the idea of sterile technique. Using sterile technique is also an issue ofhuman cleanliness and safety.Use of the PipetBefore beginning the laboratory sessions, point out the graduations on the pipette tothe students. The 100 µl and 250 µl and 1 ml marks will be used as units ofmeasurement throughout the labs.1 ml1ml750 µl500 µl250 µl100 µl6
Decontamination and DisposalIf an autoclave is not available, all solutions and components (loops and pipets) that havecome in contact with bacteria can be placed in a fresh 10% bleach solution for at least 20 min forsterilization. A shallow pan of this solution should be placed at every lab station. No matter whatyou choose, all used loops and pipets should be collected for sterilization. Sterilize petri dishesby covering the agar with 10% bleach solution. Let it stand for 1 hr or more and then pourexcess plate liquid down the drain. Once sterilized, the agar plates can be double bagged andtreated as normal trash. Safety glasses are recommended when using bleach solutions. Pleaseconsult your local environmental health and safety regulations for proper disposal.IncubationThis guide is written to reflect the use of a 37 C incubator. The transformation experimentcan be conducted without the use of an incubator, however, the number of days required toculture colonies to the optimum size depends on the ambient temperature. Best results areobtained if starter plate colonies are fresh (24–36 hr growth) and measure about 1–1.5 mm indiameter. Refrigeration of cultured plates will significantly lower transformation efficiency.The optimum temperature for growing E. coli is 37 C (98.6 F), and lower temperatures will resultin a decreased growth rate. At 28 C (82 F), two days of incubation time are required to obtainoptimum colony size. At 21 C (70 F), three days of incubation time are required to obtain optimum colony size. Adjust the advance preparation lead times and laboratory schedule according to your incubation temperature.Experimental Points – Optimizing Your pGLO Lab ExperimentPracticing TechniquesSome educators like to do a dry run of the procedures to explain sterile technique, practiceusing the pipets and loops, and practice streaking and spreading bacteria on the agar’s surface.You will have to decide what is best for your students based upon their laboratory experienceand familiarity with these techniques. Paying close attention to the following experimentalpoints will result in successful transformations.Preparing E. coli Starter (Agar) PlatesBest results are obtained when using a flask instead of a beaker to prepare the agar.Ensure that the agar is completely dissolved as uneven mixing can result in agar that will notsolidify. Follow the directions in "Advance Preparation Step 1" closely to minimize introducingcontaminants.After the starter plates are streaked with E. coli and incubated at 37 C, they should beused within 24–36 hr. Delay beyond 36 hr will compromise transformation.Transferring Bacterial Colonies from Agar Plates to MicrotubesThe process of scraping a single colony off the starter plate leads to the temptation toget more cells than needed. A single colony that is approximately 1 mm in diameter containsmillions of bacterial cells. To increase transformation efficiency, students should select 2–4"fat" colonies that are 1–1.5 mm in diameter. Selecting more than 4 colonies may decreasetransformation efficiency. Select individual colonies rather than a swab of bacteria from thedense portion of the plate; the bacteria must be actively growing for transformation to besuccessful.7
DNA TransferThe transfer of plasmid DNA from its stock tube to the transformation suspension is crucial.Students must look carefully at the loop to see if there is a film of plasmid solution across thering. This is similar to seeing a soapy film across a wire ring for blowing soap bubbles.Optionally, the students may pipet 10 µl of the pGLO plasmid into the tube labelled " pGLO"and mix well.Heat ShockThe procedure used to increase the bacterial uptake of foreign DNA is called heat shock.It is important that students follow the directions regarding time. Also important is the rapidtemperature change and the duration of the heat shock. For optimal results, the tubes containingthe cell suspensions must be taken directly from ice, placed into the water bath at 42 C for50 sec and returned immediately to the ice. The absence of the heat shock will result in a 10fold decrease in transformants; 90 sec of heat shock will give about half as many transformantsas will 50 sec of heat shock. In both cases, the experiment will work, but it has been optimized at50 sec of heat shock. Be sure to use crushed ice, instead of cubed ice for maximum transformation efficiency. Use two thermometers to check the temperature of the incubator to ensure accuracy.Spreading Transformants and ControlsDelivering an excess of transformed culture to the plates with the disposable transferpipet is counterproductive because the plates may not absorb the additional liquid andspreading will be uneven. Transferring bacterial suspensions from the microtubes to thepetri dishes requires some care. The bacteria will settle to the bottom, so the students canhold the top of a closed tube between the index finger and thumb of one hand and flick thebottom of the tube with the index finger of the other hand. Be sure that students tap thetube with their finger or stir the suspension with the pipet before drawing it up. Also, makesure that the students cover the petri dishes with lids immediately after pipetting in the transformation culture and spreading the cells. Spread the suspensions evenly around the surfaceof the agar by quickly skating the flat surface of a new sterile loop back and forth across theplate surface.Green Fluorescent Protein (GFP) Chromatography KitIf you plan to follow the pGLO bacterial transformation experiment with the GFPpurification kit (catalog #166-0005EDU), you must save the pGLO-transformed bacteriagrown on the LB/amp/ara plates. The best way to save the plates is to store them mediaside up in a cool place, such as a refrigerator. This will keep the cells alive but limit theiractive growth until you need them to start the next experiment. Storing the plates upsidedown prevents condensed moisture from smearing the colonies on the media.Ideally, plates should be used within 2–4 weeks. For longer storage, make sure that theplates are wrapped with Parafilm to prevent moisture loss.Conceptual PointsMediaThe liquid and solid nutrient media referred to as LB (named after Luria and Bertani)nutrient broth and LB nutrient agar are made from an extract of yeast and an enzymaticdigest of meat byproducts, which provide a mixture of carbohydrates, amino acids,nucleotides, salts, and vitamins, as nutrients for bacterial growth. Agar, is derived from seaweed. It melts when heated, forms a solid gel when cooled (analogous to Jello-O), and functions to provide a solid support on which to culture bacteria.8
Antibiotic SelectionThe pGLO plasmid, which contains the GFP gene, also contains the gene for beta-lactamase,which provides resistance to the antibiotic ampicillin, a member of the penicillin family. Thebeta-lactamase protein is produced and secreted by bacteria that contain the plasmid.Beta-lactamase inactivates the ampicillin present in the LB nutrient agar to allow bacterialgrowth. Only transformed bacteria that contain the plasmid and express beta-lactamase cangrow on plates that contain ampicillin. Only a very small percentage of the cells take up theplasmid DNA and are transformed. Untransformed cells cannot grow on the ampicillin selectionplates.Transformation SolutionIt is postulated that the Ca2 cation of the transformation solution (50 mM CaCl2, pH 6.1)neutralizes the repulsive negative charges of the phosphate backbone of the DNA and thephospholipids of the cell membrane to allow the DNA to enter the cells.Heat ShockThe heat shock increases the permeability of the cell membrane to D
Sep 14, 1990 · pGLO Bacterial Transformation Kit Catalog #166-0003EDU explorer.bio-rad.com For technical support call your local Bio-Rad office, or in the U.S., call 1-800-424-6723 pGLO araC GFP bla ori See individual components for storage temperature. Duplication of any p
Angular Motion A. 17 rad/s2 B. 14 rad/s2 C. 20 rad/s2 D. 23 rad/s2 E. 13 rad/s2 qAt t 0, a wheel rotating about a fixed axis at a constant angular acceleration has an angular velocity of 2.0 rad/s. Two seconds later it has turned through 5.0 complete revolutions. Find the angular acceleration of this wheel? A.17 rad/s2 B.14 rad/s2 C.20 rad/s2 .
DB-RAD 500 100-500 FtLb or DB-RAD 675 135-675 Nm DB-RAD 1000 250-1000 FtLb or DB-RAD 1350 340-1350 Nm DB-RAD 1500 375-1500 FtLb or DB-RAD 2000 510-2000 Nm Table 1.2.1: Torque Ranges 1.2.2 Battery Specifications Ensure that all Battery Specifications are followed when utilizing the Digital B-RAD Tool System. Battery Output
Please visit explorer.bio-rad.com to access our selection of language translations for Biotechnology Explorer Kit curricula. For technical service, call your local bio-rad offi ce, or in the U.S., call 1-800-4BIORAD (1-800-424-6723)
Instruction Manual Catalog #166-5015EDU explorer.bio-rad.com For technical support call your local Bio-Rad office or in the U.S. call 1-800-424-6723. PCR Fragment PCR Fragment This kit is shipped on blue ice. Open immediately upon arrival and store reagents bags at –20 C. Duplication of any part of this document is permitted for classroom .
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emit x: p90 511.6 nm rad, xn: p90 8.5%, xpn: p90 8.3%, growth projected emit x 3.6 nm rad old 1.9 nm rad emit y: p90 5.5 nm rad, yn: p90 7.9%, ypn: p90 6.5%, growth projected emit y 0.4 nm rad old 0.3 nm rad with single bunch wakes, 10% rms jitter: emit x: p90 511.6 nm rad, xn: p90 8.8%, xpn: p90 9.3% .
For technical support call your local Bio-Rad office, or in the U.S., call 1-800-424-6723 Biotechnology ExplorerTM Green Fluorescent Protein (GFP) Purification Kit Instruction Manual Catalog #166-0005EDU explorer.bio-rad.com Duplication of any p
ACCOUNTING 0452/12 Paper 1 October/November 2019 1 hour 45 minutes Candidates answer on the Question Paper. No Additional Materials are required. READ THESE INSTRUCTIONS FIRST Write your centre number, candidate number and name on all the work you hand in. Write in dark blue or black pen. You may use an HB pencil for any diagrams or graphs. Do not use staples, paper clips, glue or correction .
