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Finnpipette Finntip Good Laboratory Pipetting Guide

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Finnpipette Finntip Good Laboratory Pipetting Guide Thermo Scientific Finnpipette Finntip Good Laboratory Pipetting Guide Good Laboratory Pipetting Guide Contents Introduction 3 Over 35 years of innovation 3 Pipettes and Pipetting 4 How pipettes work 4 Pipetting terminology 5 Recommendations for pipetting different compounds 6 Getting started 6 Pipetting techniques 6 Pipetting in different applications 8 Selecting the tip 9 Ensuring optimum performance 9 Factors affecting the accuracy 10 Pipetting ergonomics 11 Decontamination Guidelines 12 Preventing cross-contamination 14 Maintenance and Service 14 Calibration of pipettes 15 Technical Information 18 Chemical resistance of plastics 18 Finntip compatibility table 19 Conversion table 20 ISO 8655 error limits 21 Warranty registration 23 Troubleshooting 23 2 Good Laboratory Pipetting Guide Good Laboratory Pipetting Guide Over 35 Years of Innovation A leader in pipetting For over 35 years, our goal has been to help customers in research and clinical laboratories to improve the speed, accuracy and precision of their pipetting. Since the introduction of the first continuously variable micropipette in 1971, more than three million Thermo Scientific Finnpipettes have been sold in 150 countries. During this time, we have listened to customer feedback to develop a wide variety of innovative models that are increasingly easy and comfortable to use. This guide outlines pipetting techniques and other practical information to help you achieve the best possible performance from your Finnpipette®. www.thermoscientific.com/finnpipette 3 Pipettes and Pipetting How pipettes work? There are two types of pipettes: air displacement and positive displacement pipettes. Air displacement pipettes are meant for general use with aqueous solutions. Positive displacement pipettes are used for highly viscous and volatile liquids. Both pipette types have a piston that moves in a cylinder or capillary. In air displacement pipettes, a certain volume of air remains between the piston and the liquid. In positive displacement pipetting, the piston is in direct contact with the liquid. Air displacement pipetting How an air displacement pipette works? Air displacement pipetting is highly accurate for standard pipetting 1. The piston moves to the This creates a partial vacuum, and applications. However, conditions, appropriate position when the the specified volume of liquid is such as atmospheric pressure as volume is set. aspirated into the tip. well as the specific gravity and viscosity of the solution, may 2. When the operating button is 4. When the operating button is effect the performance of air pressed to the first stop, the piston pressed to the first stop again, the displacement pipettes. expels the same volume of air as air dispenses the liquid. To empty indicated on the volume setting. the tip completely, the operating button is pressed to the second 3. After immersing the tip into stop (blow-out). the liquid, the operating button is released. piston pipette tip Aspirating the liquid (steps 1-3) Dispensing the liquid (step 4) 4 Pipettes and Pipetting Positive displacement pipetting How the Thermo Scientific Finnpipette Stepper works. The Thermo Scientific Finnpipette Stepper repeater pipette uses the 50 ml 50 1. The piston inside the tip rises ® positive displacement principle. when filling the tip with liquid. Finntip Disposable Stepper microsyringe tips have a piston inside a cylinder 2. When the dispensing lever is unit. This helps to avoid sample-to- pressed down, the piston descends ml 1=1 sample cross-contamination (also and the selected volume is known as sample carry-over) and dispensed. The dispensing lever contamination due to the aerosol has to be pressed once for each effect. dispensing stroke (= step). Pipetting terminology The following terms are used throughout this guide. Aspirate – to draw the liquid up into the pipette tip Dispense – to discharge the liquid from the tip Blow-out – to discharge the residual liquid from the tip Calibration check – to check the difference between the dispensed liquid and the selected volume Adjustment – altering the pipette settings so that the dispensed volume is within the specifications Recommendations for pipetting different compounds Solution/ Examples Pipette Tip Pipetting Comments compound tech- nique Aqueous Buffers, diluted salt Air displacement Standard Forward solution solutions Viscous Protein and nucleic Air displacement Standard or wide orfice Reverse Pipette slowly to avoid bubble formation. solution acid solutions, glycerol, Tween 20/40/60/80 Positive displacement Positive displacement Volatile Methanol, hexane Air displacement Filter Reverse Pipette rapidly to avoid evaporation. Carbon compounds filter tips prevent vapor going into the pipette Positive displacement Positive displacement very effectively Body fluids Whole blood, serum Air displacement Standard or wide Pipetting Residual liquid can be found on the outer orifice tip of hetero- surface of the tip. Wipe the tip against the geneous edge of the vessel to remove this liquid before samples dispensing. Nucleotide Gonomic DNA, PCR Air displacement Filter or wide orifice Forward For genomic DNA wide orifice tips can be solutions products used to eliminate mechanical shearing. Positive displacement Positive displacement Radioactive 14Carbonate, 3H-thy- Air displacement Filter Forward compounds midine Positive displacement Positive displacement Acids/alkalis H SO , HCI, NaOH Air displacement Filter Forward 2 4 Toxic Air displacement Filter Forward or samples reverse Positive displacement Positive displacement www.thermoscientific.com/finnpipette 5 Pipettes and Pipetting Getting started • Check your pipette at the • Tips are designed for single • Pipette parallel samples in a beginning of your working day use. They should not be similar way. for dust and dirt on the outside. cleaned for reuse, as their If needed, wipe with 70% metrological characteristics will • Avoid turning the pipette on ethanol. no longer be reliable. its side when there is liquid in the tip. Liquid might get into • Check that you are using • Pre-rinsing (three to five times) the interior of the pipette and tips recommended by the the tip with the liquid to be contaminate the pipette. manufacturer. To ensure pipetted improves accuracy. accuracy, use only high-quality This is especially important • Avoid contamination to or from tips made from contamination- when pipetting volatile hands by using the tip ejector. free polypropylene. compounds since it prevents liquid from dripping out of the • Always store pipettes in an tip. upright position when not in use. Finnpipette stands are ideal for this purpose. Pipetting techniques Forward pipetting Ready position 1 2 3 4 Otherwise no: Formation of bubbles 3. Dispense the liquid into the First stop or foam in the tip or in the test tube receiving vessel by gently pressing Second stop or well. the operating button to the first stop. After one second, press the operating Yes: When pipetting and mixing a 1. Press the operating button to the button to the second stop. This action sample or reagent into another liquid. first stop. will empty the tip. Remove the tip from the vessel, sliding it along the 2. Dip the tip into the solution to a The forward technique is wall of the vessel. recommended for aqueous solutions, depth of 1 cm, and slowly release such as buffers, diluted acids or the operating button. Withdraw 4. Release the operating button to the alkalis. the tip from the liquid, touching it ready position. against the edge of the reservoir to remove excess liquid. 6 Pipettes and Pipetting Pipettes and Pipetting Repetitive pipetting Ready position 1 2 3 4 1. Press the operating button to the 3. Dispense the liquid into the second stop. receiving vessel by gently pressing First stop the operating button to the first Second stop 2. Dip the tip into the solution to a stop. Hold the button in this depth of 1 cm, and slowly release position. Some liquid will remain in the operating button. Withdraw Yes: Especially for adding reagents the tip, and this should not be dis- the tip from the liquid, touching it into tubes or into the wells of pensed. against the edge of the reservoir to microplates. remove excess liquid. 4. Continue pipetting by repeating This technique is intended for steps 2 and 3. repeated pipetting of the same volume. Reverse pipetting viscosity or a tendency to foam. 3. Dispense the liquid into the Ready position 1 2 3 4 5 This method is also recommended receiving vessel by depressing the First stop for dispensing small volumes. It can operating button gently and steadily Second stop also be used with air displacement to the first stop. Hold the button pipettes. in this position. Some liquid will Yes: For pipetting samples or 1. Press the operating button to the remain in the tip, and this should reagents when no mixing into second stop. not be dispensed. another liquid is required. 2. Dip the tip into the solution to a 4. The liquid remaining in the tip Reverse pipetting avoids the risk depth of 1 cm, and slowly release can be pipetted back into the of splashing, and foam or bubble the operating button. This action will original solution or thrown away formation. fill the tip. Withdraw the tip from with the tip. the liquid, touching it against the The reverse technique is used for 5. Release the operating button to edge of the reservoir to remove pipetting solutions with a high the ready position. excess liquid. Pipetting of heterogeneous samples 1. 3. + 4. Press the operating button Ready position 1 2 3 4 5 6 Press the operating button to the first stop. Dip the tip into to the first stop and release it First stop the sample. Make sure the tip is slowly to the ready position. Do not Second stop sufficiently below the surface. remove the tip from the solution. Repeat this process until the 2. Release the operating button interior wall of the tip is clear. Yes: When prerinsing the tip is not slowly to the ready position. This possible and the full sample should action will fill the tip with the 5.
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