Thermolab Sartorius MC5 Manual
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Thermolab Sartorius MC5 Manual

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Liquid Handling
Application Notebook
Tips on how to
pipette

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Summary of Contents for Thermolab Sartorius MC5

  • Page 1 Liquid Handling Application Notebook Tips on how to pipette...

  • Page 3: Table Of Contents

    Contents Pipetting terms ......................4 Types of pipettes ..................... 4 General guidelines and pipetting techniques ............7 Recommendations for pipetting different compounds ......... 10 Pipetting guidelines for selected compounds ............11 Preventing cross-contamination ................14 Finntip Filter tests ....................15 The plastics of various Finntips ................

  • Page 4: Pipetting Terms

    Pipetting terms Aspirate - to draw up the sample Dispense - to deliver the sample Blow-out - to empty the tip completely Calibration check - checking the difference between the dispensed volume and the selected volume Adjustment - altering the pipette so that the dispensed volume is within the specifications Types of pipettes Although Thermo Labsystems supplies pipettes for all application needs, most lab-...
  • Page 5 How does an air displacement pipette work? 1. The piston moves to the appropriate position when the volume is set. 2. When the operating button is pressed to the first stop, the piston expels the same volume of air as indicated on the volume setting. 3.
  • Page 6 Positive displacement pipetting Positive displacement pipetting is used for applications like PCR and other DNA amplification techniques. The microsyringe tips used in positive displacement pi- pettes are disposable. This helps to avoid sample-to-sample cross-contamination (also known as sample carry-over), and contamination due to the aerosol effect. •...

  • Page 7: General Guidelines And Pipetting Techniques

    General guidelines and pipetting techniques General guidelines Check your pipette at the beginning of your working day for dust and dirt on the outside. If needed, wipe with 70% ethanol. Set the volume within the range specified for the pipette. Hold the pipette so the ‘grippy finger rest’...
  • Page 8 Forward pipetting 1. Press the operating button to the first stop. 2. Dip the tip into the solution to a depth of 1 cm, and slowly release the operating button. Wait 1-2 seconds and withdraw the tip from the liquid, touching it against the edge of the reservoir to remove excess liquid.
  • Page 9 Repetitive pipetting This technique is intended for repeated pipetting of the same volume. 1. Press the operating button to the second stop. 2. Dip the tip into the solution to a depth of 1 cm, and slowly release the operating button. Withdraw the tip from the liquid, touching it against the edge of the reservoir to remove excess liquid.

  • Page 10: Recommendations For Pipetting Different Compounds

    Recommendations for pipetting different compounds...

  • Page 11: Pipetting Guidelines For Selected Compounds

    Pipetting guidelines for selected compounds Body Fluids Whole Blood Pipette + tip combination: Choose an air displacement pipette and a standard or wide orifice tip. Technique: Use the whole blood pipetting technique. Reverse pipetting should be used if high accuracy is needed. Notice: Some blood can remain in the tip and on the outer surface.
  • Page 12 vessel to remove excess liquid outside the tip before dispensing. The use of a pos- itive displacement pipette and tip is also useful for pipetting glycerol. Tween 20, 10% solution Pipette + tip combination: Choose an air displacement pipette and a standard or wide orifice tip. Technique: Use the reverse pipetting technique.
  • Page 13 Concentrated acids and bases Pipette + tip combination: Choose an air displacement pipette and a filter tip. Technique: Use the forward pipetting technique. NaOH Pipette + tip combination: Choose an air displacement pipette and a filter tip. Technique: Use the forward pipetting technique. Notice: Some acids or bases vaporise easily (e.g.

  • Page 14: Preventing Cross-Contamination

    Notice: 1. To get accurate results, calibrate the pipette with the volatile compound you want to pipette. If you use air displacement pipettes, aspirate and dispense the liquid a few times keeping the tip in the liquid. By doing so, the air inside the pipette will be saturated with vapour of the volatile compound.

  • Page 15: Finntip Filter Tests

    Finntip Filter tests Acid test • 35% and 5% trifluoroacetic acid (TFA) was used as test liquid. • Finntip Filters prevented acids vapours of a 35% TFA solution of coming through the filter. • When pipetting a 5% solution of TFA using a standard tip, no vapour came into the interior of the pipette.

  • Page 16: Troubleshooting

    Troubleshooting Finnpipette ® Digital Problem Possible cause Solution Leakage Tip incorrectly attached. Attach tips firmly, keeping pipette vertical and pressing pipette evenly to rack. Do not bang. Wrong tip size or shape. Check that the size and shape are correct. Pipette incorrectly assembled after Check the assembly according taking apart.

  • Page 17: Calibrating Your Pipettes

    Calibrating your pipettes Calibration of Pipettes Calibration of pipettes officially means determining the difference between the dis- pensed volume and the selected volume. Adjustment means altering the pipette so that the dispensed volume is within certain specifications. All Finnpipettes are factory calibrated and adjusted to give the volumes as spec- ified with distilled or deionised water.
  • Page 18 The scale graduation value of the balance should be chosen according to the se- lected pipette volume. Volume range Readable graduation Sartorius model (example) Under 10 µl 0.001 mg Sartorius MC5 10 - 100 µl 0.01 mg Sartorius MC210 Above 100 µl 0.1 mg Sartorius MC210 Note: Check the calibration of your balance regularly using known weights.
  • Page 19 Manual multichannel pipettes The pipette is held in the calibration room for at least 2 hours before calibration to reach equilibrium with the test room conditions. The pipette is checked at the maximum volume (nominal volume) and at the minimum volume or 10% of the maximum volume, whichever is higher. For example, Finnpipette 0.5 - 10 µl is tested at 10 µl and 1 µl.
  • Page 20 calibration nut at the top of the handle. Turn the service tool clockwise to increase the volume or counter clockwise to decrease the volume. After the adjustment, check the calibration as described above. Manual multichannel pipettes The adjustment is done at the lower volume with one of the middle channels. Place the service tool that comes with the pipette into the openings of the calibration nut at the top of the handle.

  • Page 21: Formulas For Calculating Results

    factors have different values depending on the module, so please check that you choose the right module. 5. Enter these factors to the OLD FACTORS fields of the calibration software. 6. To get the new factors, click the CALCULATE button. The new factors will be displayed.
  • Page 22 Precision (random error) Precision refers to the repeatability of the pipettings. It is expressed as standard deviation (s) or coefficient of variation (cv). In addition to the features of the pi- pette, laboratory practice and user experience are the main factors affecting preci- sion.

  • Page 23: Conversion Tables

    Conversion tables Table 1: Values of the conversion factor Z (µl mg ) as a function of temperature and air pressure, for distilled water. Temperature °C Air pressure hPA (mbar) 1013 1067 1.0018 1.0018 1.0019 1.0019 1.0020 1.0020 15.5 1.0018 1.0018 1.0019 1.0020...

  • Page 24: Ensuring Optimum Performance

    Ensuring optimum performance Error-free pipetting requires both precision and accuracy. A number of factors can affect these specifications, which are the main quantitative parameters for evaluat- ing pipette performance. What are accuracy and precision? For example when the set volume is 20 µl: 20 µl 20 µl 20 µl...
  • Page 25 The effect of the pipetting position (e.g. using a 2-10 ml pipette) Accuracy Accuracy 0.2-0.4% 0.6-0.8% Accuracy 1-1.2% 1 cm 3 cm 3-4 cm 1. Pipette vertical, tip immersed about 1 cm into the liquid. 2. Pipette vertical, tip immersed about 3 cm into the liquid. 3.

  • Page 26: Factors Affecting The Accuracy Of Air Displacement Pipettes

    Factors affecting the accuracy of air displacement pipettes Temperature The most important factor in pipetting accuracy is the liquid temperature. The fig- ure below shows the change in volume when the liquid has a different tempera- ture than the pipette and air. If the temperature of the liquid, pipette and air is the same, the accuracy is not significantly affected.

  • Page 27: Maintenance Of Your Finnpipette

    Maintenance of your Finnpipette Finnpipettes are easy to service in the laboratory using the tools provided with the pipette. All Finnpipettes come with detailed instructions on how to disassemble the pipette. Instructions for routine in-lab maintenance are also included. Short-term checking: At the beginning of each workday, the pipette should be checked for dust and dirt on the outside surfaces.

  • Page 28: General Guidelines For Decontaminating Pipettes When Working With Different Liquids

    General guidelines for decontaminating Pipettes when Working with Different Liquids Liquid Handling, Special features Decontamination Aqueous Pipettes are calibrated with Open pipette, rinse contaminated parts solutions and distilled water. Results are well with distilled water, and allow to dry buffers extremely accurate.

  • Page 29: Chemical Compatibility Of Plastics

    Chemical compatibility of plastics These are general guidelines, not performance guarantees. Factors such as concentration, temperature and length of exposure can affect performance. Finntip Plungers Tip cones Tip cones BioMate BioMate (Stepper, (BioControl, Finnpipette, (Finnpipette MCP (adaptor) (nose cone) Finntip PDP) Digital MCP , SCP) Classic, MCP Colour) Chemical Class...
  • Page 30 These are general guidelines, not performance guarantees. Factors such as concentration, temperature and length of exposure can affect performance. Finntip Plungers Tip cones Tip cones BioMate BioMate (Stepper, (BioControl, Finnpipette, (Finnpipette MCP (adaptor) (nose cone) Finntip PDP) Digital MCP , SCP) Classic, MCP Colour) Chemical Class Polypro-...

  • Page 31: Frequently Asked Questions

    Frequently asked questions Finnpipette Digital Question: How can I sterilize the pipette? Answer: Finnpipette Digital is fully autoclavable (121°C, 20 min). The pipette can also be exposed to UV radiation. The colour of the handle may turn yellowish after pro- longed exposure.
  • Page 32 and then push the + and - buttons as well. CALIBRATE text is now blinking, push SET to accept. The current HK-factor is now blinking followed by the PK-factor. Note: These factors have different values depending on the module, so please check that you choose the right module.
  • Page 33 Question: How can I pipette viscous liquids? Answer: You can do so using an air displacement pipette with standard or wide orifice tip (reverse pipetting, slowly). An alternative to this is to use a positive displacement system. Question: How can I prevent liquid dropping out of the tip when pipetting volatile com- pounds? Answer: If you use air displacement pipettes, aspirate and dispense the liquid a few times...

  • Page 34: Making Your Lab Work Lighter

    Making your lab work lighter High quality lab work needs good ergonomics. Work goes smoother and does not cause too much stress. Nowadays a lot of attention is paid to the ergonomics of lab equipment and furniture, but it is also important to do things in the right way. Here you find some examples of how to improve your way of working (with the kind permission of the Centre for Occupational Safety, Finland).

  • Page 35: Finnpipette Warranty Policy

    Make repetitive work lighter: * Use both hands: use your left hand (if you are right handed) when using a vortex mixer, an electronic pipetting aid or the computer mouse. * Avoid unnecessary squeezing of lab tools (e.g. pipettes). * Choose lighter or electronic pipettes and dispensers. * Use tubes that can be opened and closed easily.
  • Page 36 Sorvaajankatu 15 P .O. Box 208 FIN-00811 Helsinki, Finland Tel. +358-9-329 100 Fax +358-9-3291 0415 www.labsystems.fi A Thermo Electron Business...

This manual is also suitable for:

Sartorius mc210

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