oSpectrometer® nual basic gEN) manual Register your instrument! www.eppendorf.
Copyright © 2014 Eppendorf AG, Hamburg. No part of this publication may be reproduced without the prior permission of the copyright owner. Trademarks Eppendorf® and the Eppendorf logo, Eppendorf BioSpectrometer®, Eppendorf SpectraZoom,® and UVette® are registered trademarks of Eppendorf AG, Hamburg, Germany. Cy® is a registered trademark of GE Healthcare UK Ltd., Buckinghamshire, UK. Hellma® is a registered trademark of Hellma GmbH & Co. KG, Müllheim, Germany.
Table of contents Eppendorf BioSpectrometer® basic English (EN) Table of contents 1 Operating instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 1.1 Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 1.2 Danger symbols and danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Table of contents Eppendorf BioSpectrometer® basic English (EN) 6 Methods. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 6.1 Selecting a method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 6.2 Photometry method description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 6.2.
Table of contents Eppendorf BioSpectrometer® basic English (EN) 11 Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 11.1 Power supply. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 11.2 Ambient conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Table of contents Eppendorf BioSpectrometer® basic English (EN)
Operating instructions Eppendorf BioSpectrometer® basic English (EN) 1 Operating instructions 1.1 Using this manual Read this operating manual completely before using the device for the first time. Also observe the instructions for use of the accessories. This operating manual is part of the product. Thus, it must always be easily accessible. Enclose this operating manual when transferring the device to third parties.
Operating instructions Eppendorf BioSpectrometer® basic English (EN) 1.3 Symbols used Depiction Meaning 1. 2. Actions in the specified order Actions without a specified order • List Press this key to perform the described action. or sample Press this softkey to perform the described action.
Operating instructions Eppendorf BioSpectrometer® basic English (EN) 1.
Operating instructions Eppendorf BioSpectrometer® basic English (EN)
Product description Eppendorf BioSpectrometer® basic English (EN) 2 Product description 2.1 Main illustration Abb. 2-1: Front and rear view 3 2 3 ce absorban absorban 1 ce height 8.5 m m 1 2 3 abc 4 def 5 gh i jkl 7 pq rs 8 tuv meth od 6 mno 9 wxyz exit func tion 0 del ete µ % ente r standard blan k sample 10 Fig.
Product description Eppendorf BioSpectrometer® basic English (EN) 2.3 Features The BioSpectrometer basic is a UV/Vis spectrophotometer for measuring liquids in cuvettes in a wavelength range of 200 nm to 830 nm. It is intended for use in development and research in the fields of molecular biology, biotechnology, biochemistry and cell biology. Glass and plastic cuvettes in a volume range of 1 μL to 3000 μL can be used. 2.3.
Safety Eppendorf BioSpectrometer® basic English (EN) 3 Safety 3.1 Intended use The BioSpectrometer basic is to be used in molecular biology, biochemistry and cell biology research laboratories. The BioSpectrometer basic is exclusively intended for use indoors. All country-specific safety requirements for operating electrical equipment in the laboratory must be observed.
Safety Eppendorf BioSpectrometer® basic English (EN) WARNING! Damage due to UV radiation. Microliter cuvettes, e.g., Hellma® TrayCell (or microliter cuvettes with a similar design) divert the radiation from the light source within the cuvette so the radiation can escape upward when the lid is not closed. Before starting a measurement, ensure that the lid on the microliter cuvette is not open.
Safety Eppendorf BioSpectrometer® basic English (EN) NOTICE! Damage to electronic components due to condensation. Condensate can form in the device after it has been moved from a cool environment to a warmer environment. After installing the device, wait at least for 3 h. Only then connect the device to the mains. NOTICE! Function impairment due to mechanical damage.
Safety Eppendorf BioSpectrometer® basic English (EN) 3.5 Safety instructions located on the device Depiction Meaning Location Hazard point Rear side of the device Follow the operating manual. Gerät nach dem Öffnen justieren! Adjust device after opening! The device needs to be readjusted after it has been opened. Do not open the device.
Installation Eppendorf BioSpectrometer® basic English (EN) 4 Installation 4.1 Preparing installation Keep the transport carton and the packing material for subsequent safe transport or storage. Check the completeness of the delivery using the information in the delivery package (see Delivery package on p. 11). Check all parts for any transport damage. 4.
Installation Eppendorf BioSpectrometer® basic English (EN) 4.4.2 Thermal printer DPU-414 Connect the thermal printer DPU-414 to the serial printer connection. 1. Connect the printer cable to the serial printer connection 9 and tighten the locking screws.(see Main illustration on p. 11). 2. Connect the printer cable to the printer and tighten the locking screws as well. 3.
Installation Eppendorf BioSpectrometer® basic English (EN) 4.5 Connecting PC or USB stick for data export You can connect a FAT 32-formatted USB stick to the USB port 4 (see Main illustration on p. 11). Alternatively, you can connect the device for the data export directly to a PC by using a USB cable: Prerequisites • PC with Windows, version XP, SP2 or higher version. • USB cable with a type A and type B plug each.
Installation Eppendorf BioSpectrometer® basic English (EN)
Operation Eppendorf BioSpectrometer® basic English (EN) 5 Operation 5.1 Overview of operating controls Abb. 5-1: Control panel of the BioSpectrometer basic Fig.
Operation Eppendorf BioSpectrometer® basic English (EN) Key: Function Keypad: Enter digits and text. Keys 1 to 9 as well as 0: When entering text, next to numbers you also can enter letters and special characters by pressing the key several times. Alternatively, you can switch to a displayed keyboard with the [Keyboard] key. Outside of entry fields: Call up method selection. Outside of entry fields: Call up function selection. Softkey: Select functions.
Operation Eppendorf BioSpectrometer® basic English (EN) 5.1.1 Entering text You can enter texts when assigning method names and result units. Restriction: Only digits, letters and the underscore "_" are allowed for method names. Entry via keyboard: Use the and cursor keys to navigate within the entry field and to change single positions in the name. Softkeys: • [Keyboard]: Display keyboard. • [abc]: Change between upper and lower case letters when making entries with the keypad.
Operation Eppendorf BioSpectrometer® basic English (EN) 5.2 Inserting the cuvette Standard rectangular glass or plastic cuvettes can be inserted in the cuvette shaft: • External dimensions: 12.5 mm × 12.5 mm • Height of light path: 8.5 mm higher than cuvette base • Total height: min. 36 mm The cuvettes must be optically transparent for the respective measuring wavelength.
Operation Eppendorf BioSpectrometer® basic English (EN) 5.3 Summary of the measuring procedure 5.3.1 Preparing the measurement 1. Switch on the device and, if required, the printer. The device performs a self test (taking approx. 1 minute) and displays the method selection. 2. Make ready the cuvettes for the measurements (see Inserting the cuvette on p. 24). 3. Prepare the measuring solutions for measuring the blank values, if required, also the standards and the samples. 4.
Operation Eppendorf BioSpectrometer® basic English (EN) 5.3.2.2 Checking parameters Check the parameter setting. The [Page dn] and [Page up] softkeys allow you to call up the parameter list pages. You can modify and save parameters using [Edit]. 5.3.2.3 Measuring the blank and standards For evaluations without standards (e.g. DNA measurements), this method step is omitted. 1. Start by measuring a blank (blank key). 2. Then measure all standards one by one (standard key).
Operation Eppendorf BioSpectrometer® basic English (EN) 5.3.2.4 Measuring samples The sample key is used for measuring your samples consecutively. Blank results will remain saved for the duration of one series of measurement. However, a new blank measurement always is possible. (The adjacent figure shows a measuring procedure with evaluation via the standard curve and, in addition to the sample result, the graph of the standard evaluation.) 5.3.2.5 Finalizing the method 1.
Operation Eppendorf BioSpectrometer® basic English (EN) 5.3.2.7 Printing and exporting 1. Compose data packets for all samples or for selected samples. 2. Print the data, save them to a USB stick or transfer them to a PC via a USB cable.
Operation Eppendorf BioSpectrometer® basic English (EN) 5.3.3 Important measurement instructions Check for each measurement: • For plastic cuvettes: How many consecutive measurements can be reliably carried out in the cuvette? • Measure the cuvette blank value before the sample or standard measurements in order to compensate the cuvette blank value in addition to the reagent blank.
Operation Eppendorf BioSpectrometer® basic English (EN)
Methods Eppendorf BioSpectrometer® basic English (EN) 6 Methods 6.1 Selecting a method Methods and method templates are delivered preprogrammed. The methods are organized in main groups and subgroups. Write-protected methods The most important methods in molecular biology. Parameters can be modified, but the modified parameters must be saved under a new method name. Non-write-protected methods You can change parameters any number of times and start the measurement right after saving.
Methods Eppendorf BioSpectrometer® basic English (EN) Tab. 6-2: Softkeys in method selection [Cut] and [Paste] Cut and paste methods. [Copy] and [Paste] Copy and paste methods. [Delete] Delete methods. [Rename] Rename methods. Copied or cut methods can be added to a different folder under Favorites, or added to the original folder under a new name. Use the cursor keys to navigate to the Methods column of the desired folder and press [paste] for adding the method. 6.
Methods Eppendorf BioSpectrometer® basic English (EN) 6.2.2 Routine method group The methods for the Routine group are preprogrammed as fixed methods. Therefore, a new method name is required after the method parameters in the fixed preprogrammed methods have been modified. Nucleic acids • Determination of the concentration of nucleic acids through measurement at 260 nm and evaluation via factor. • Various nucleic acid methods, such as dsDNA or RNA, are preprogrammed.
Methods Eppendorf BioSpectrometer® basic English (EN) Dye labels • For dye-labeled biomolecules: Concentration determination of the biomolecule (nucleic acid or protein) via measurement at 260 or 280 nm and measurement of the dye in one measuring procedure. • Evaluation with factor. In addition to the biomolecule, up to two dyes can be measured at the same time as two different wavelengths. • Additional: evaluation of the frequency of incorporation (FOI) of the dye.
Methods Eppendorf BioSpectrometer® basic English (EN) 6.2.4 Advanced method group Dual wavelength • Measurement at two wavelengths and evaluation of the measured absorbance values via two basic formulas (subtraction, division) • Variants of the basic formulas can be defined. • The result can be evaluated with a factor, with a standard or with a standard series. • Methods are preprogrammed for calculation, subtraction and division, and subsequent factor evaluation. 6.
Methods Eppendorf BioSpectrometer® basic English (EN) Parameter Entry Explanation Unit Selection: Unit for the concentration result. mg/mL | μg/mL | ng/ In the preprogrammed methods of the Routine group, the mL | pg/mL | μg/μL | selection is restricted to units that are useful for these methods. mg/dL | μmol/mL | nmol/mL | pmol/mL | pmol/μL | U | U/mL | U/L | % | Abs | A/min In addition, further units are freely programmable in the General Method Parameters/Units function. Max. 7 digits.
Methods Eppendorf BioSpectrometer® basic English (EN) Parameter Entry Explanation Factor Value input: Factor. Limit: max. of 6 digits including decimal point. Factor for converting absorbance values into the concentration. Negative factors can also be entered for the following method groups: Dual wavelength, Factor. For the Dye labels method group the factors are not entered into the method procedure individually.
Methods Eppendorf BioSpectrometer® basic English (EN) Parameter Entry Explanation Correct A260 1 Selection: on | off Only for the Dye labels method group. Correction of the influence of the dye spectrum on the absorbance with the measuring wavelength of the biomolecule (260 nm or 280 nm). Some of the dye spectra have a low absorbance at 260 and 280 nm. These absorbances distort the calculations for the nucleic acids or the proteins of these methods.
Methods Eppendorf BioSpectrometer® basic English (EN) Parameter Entry FOI Selection: Only for the Dye labels method group. none | dye/kb | pmole/ Display of the FOI in addition to the result of the sample μg measurement. The FOI (frequency of incorporation) is a measure for the number of dye molecules per nucleic acid molecule that are integrated into the nucleic acid. Units are "dye/kb" (dye molecules per 1000 bases) or "pmole/μg" (pmol dye per μg nucleic acid). "None": no FOI calculation.
Methods Eppendorf BioSpectrometer® basic English (EN) 6.4 Method procedure Wizard: the wizard at the top of the display will take you through the method procedure. The currently active method step is highlighted. A method procedure is composed of a maximum of 5 steps. The currently active step is highlighted visually. After the last step, print & export, of a measuring series, the start of a new measuring series is offered as a next step. It once again starts with the sample measurement.
Methods Eppendorf BioSpectrometer® basic English (EN) 6.4.1 Check parameters Softkeys • [Page dn] and [Page up]: Change between 1 to 3 parameter list pages. • [Edit]: Switch to the parameter edit mode. Editing mode for parameters: Modified parameters are marked with a red star until the modification has been saved. Softkeys • [Save] and [Save as]: Save changes. When using [Save as] you have to rename the method.
Methods Eppendorf BioSpectrometer® basic English (EN) 6.4.2 Measure standards The first standard to be measured is marked on the display. After the blank value (blank key) measure all standards (standard key) one by one. When measuring more than one replicate per standard, the average value for each standard is calculated and displayed automatically. By using the and cursor keys, you also can select certain standards for measurement. Individual standards can be remeasured as well.
Methods Eppendorf BioSpectrometer® basic English (EN) 6.4.3 Measure samples The sample key is used for measuring your samples consecutively. Blank results remain saved for one measuring series, but a new blank result measurement can be performed at any time. With the and keys you can navigate between the sample results that have been achieved in the measuring series up to this point. Results display: • The concentration result (6 digits with floating point) is clearly emphasized.
Methods Eppendorf BioSpectrometer® basic English (EN) Enter dilution The [Dilution] softkey is activated after the blank (blank key) has been measured. 1. Press the [Dilution] softkey. 2. Enter the volumes for the sample (up to 3 digits) and for the dilution buffer (up to 4 digits). The device will multiply the following sample results by the calculated dilution factor. Softkeys • [Clear dil.]: Delete values for sample dilution. • [OK]: Confirm sample dilution and return to sample measurement.
Methods Eppendorf BioSpectrometer® basic English (EN) Result image with dilution and ID Result image with dilution and sample ID 6.4.4 Measure samples: Results displays This section contains a display of typical results displays for all method groups and an overview of additional results data, which can be accessed using the [Data] softkey.
Methods Eppendorf BioSpectrometer® basic English (EN) Method group Results display Scan Explanation Results display: • Scan (graph with absorbance wavelength display) • Navigate between the measuring points in the graph with and . Routine main group Nucleic acids Results display: • Concentration result with absorbance at the measuring wavelength • If activated in the parameters: Ratios A260/A280 • If activated in the parameters: Scan.
Methods Eppendorf BioSpectrometer® basic English (EN) Method group Proteins direct UV Results display Explanation Results display: • Concentration result with absorbance at the measuring wavelength • If activated in the parameters: Scan. Navigate between the measuring points on the graph which are used for the result calculation with and . Additional data ([Data] softkey):. If the corresponding parameters have been activated: • Absorbance value for 260 nm.
Methods Eppendorf BioSpectrometer® basic English (EN) Method group Results display Dye labels Explanation Results display: • Concentration results with absorbance at the measuring wavelength of the biomolecule. • If activated in the parameters: Scan. Navigate between the measuring points in the graph with and . Additional data ([Data] softkey):. If the corresponding parameters have been activated: • Ratios A260/A280 and A260/A230.
Methods Eppendorf BioSpectrometer® basic English (EN) Method group Results display Explanation Advanced main group Dual wavelength Results display: • Concentration result: calculated from Acalc. with factor or standard evaluation. • Acalc.: calculated using the formula, defined in the parameters, created from the absorbances measured on both wavelengths. • Absorbance values that were measured at the two measuring wavelengths. Additional data ([Data] softkey):.
Methods Eppendorf BioSpectrometer® basic English (EN) 6.4.5 Process results The sample measurement is followed by two optional steps in the method sequence: process results and print & export. In the process results step, you can postprocess the results for some methods. Example: Changing the spectra section of a scan. As for the result display, you can navigate between the sample results of the measurement series with the and cursor keys and select results for postprocessing. Tab.
Methods Eppendorf BioSpectrometer® basic English (EN) After changes have been made, you can exit the current mode using the two softkeys at right: • [Save]: Save changes and return to the process results method step. • [Cancel]: Cancel and return to the process results method step. After the changes have been saved you can apply them to all samples of the measuring series with [Yes].
Methods Eppendorf BioSpectrometer® basic English (EN) 6.4.6 Process results: Options Zoom Press the [Zoom] softkey and select one of the following versions. Variant [spectra]: • Cursor keys and : Move the wavelength cursor. It determines the zoom center above the x-axis. • Cursor keys and : Gradually zoom in and out of the displayed section of the x-axis using the SpectraZoom procedure.
Methods Eppendorf BioSpectrometer® basic English (EN) More calculations Press the [More calc.] softkey. Nucleic acids method group: • After the molar mass has been entered (in base/ base pairs or in kDa): Convert the concentration result to the molar concentration. • After the sample volume has been entered: Calculate the total amount in the sample.
Methods Eppendorf BioSpectrometer® basic English (EN) Peak detection Press the [Peaks] softkey. For the peak detection you can alternate between two criteria: • λ grid: Evaluation grid on the wavelength scale for the peak detection (e.g., 10 nm). 10 nm example: The spectra section from -5 nm to +5 nm is evaluated in relation to the peak to be detected. • Min. Δ Abs: Minimum difference between the peak to be detected and the lowest absorbance in the evaluation grid.
Methods Eppendorf BioSpectrometer® basic English (EN) 6.4.7 Print & export In the last optional method step, you can assemble data packets for all samples of a series of measurements, or selected samples of a series of measurements, for printing to the printer, export to a USB stick or export to a PC using a USB cable. Select data packets • Use the cursor keys for navigating and confirm with enter. Softkeys • [Print]: Start printing. • [Export]: Start export. • [Sample]: Select individual sample results.
Methods Eppendorf BioSpectrometer® basic English (EN) Export to USB stick 1. Connect a FAT 32-formatted USB stick to the USB port 4 (see Main illustration on p. 11). 2. Start with [Export] "export to an external storage medium". Export to PC Requirement for the PC operating system: Windows XP, SP2 or a higher version. 1. Connect the device to the PC by using the USB cable on the USB port 8 (see Main illustration on p. 11). 2.
Methods Eppendorf BioSpectrometer® basic English (EN) Select data packets Results Primary result data; cannot be selected because they are always transferred. Data Additional results data that are displayed during the measurement using the [Data] softkey. Graph Absorbance-wavelength-spectrum. Graph data The basic numeric data for the graph. "export only": Only available for export, i.e., not for printing. Parameters Method parameters Standards/results Results data of the standard evaluation.
Methods Eppendorf BioSpectrometer® basic English (EN)
Functions Eppendorf BioSpectrometer® basic English (EN) 7 Functions 7.1 Functions of the User main group With the function key or the [Function] softkey, you reach a menu containing functions like device settings or calling up saved results. The functions are structured in 3 columns analog to the method selection. The functions in the User main group are accessible to you.
Functions Eppendorf BioSpectrometer® basic English (EN) 7.1.1 Results memory In the right column, select the method for which you would like to call up saved results. Confirm with enter. Select the desired series of measurement with the cursor keys. Confirm with enter. As in the method procedure, you can also successively switch between the display of the parameters, standards, sample results and, finally, the data packets for print and export.
Functions Eppendorf BioSpectrometer® basic English (EN) If you would like to print or export results, select the data packets. The procedure for print and export and the meaning of the function keys corresponds to the print & export method step. 7.1.2 General method parameters In the right column, select the parameter group for which you would like to edit parameters. Confirm with enter.
Functions Eppendorf BioSpectrometer® basic English (EN) To edit a parameter group, use and to select the parameter which you would like to edit. Confirm with enter. Softkeys • [OK]: Save entry and return to the parameter group selection. • [Cancel]: Return to the parameter group selection without making any changes.
Functions Eppendorf BioSpectrometer® basic English (EN) Tab. 7-2: Parameter in General Method Parameter Parameter Explanation Proteins These parameters are loaded into the method parameters when a protein is selected during the programming of a method in the Dye labels and Proteins direct UV groups. • Protein name • Factor • A0.1% • Ext.coeff.
Functions Eppendorf BioSpectrometer® basic English (EN) 7.1.3 Absorbance spectra library In the right column, you select the spectrum which you would like to call up and confirm with enter. Softkeys • [Export] and [Print]: Export to a USB stick or print to a PC using a USB cable (see Print & export on p. 55). • [OK]: Return to the function selection. 7.1.4 Device settings The following settings can be modified: • Language: German, English, French, Spanish, Italian. • Date and time.
Functions Eppendorf BioSpectrometer® basic English (EN) 7.1.6 Info The Copyright menu item contains license information on the Open Source software.
Functions Eppendorf BioSpectrometer® basic English (EN)
Maintenance Eppendorf BioSpectrometer® basic English (EN) 8 Maintenance 8.1 Cleaning DANGER! Electric shock as a result of penetration of liquid. Switch off the device and disconnect the power plug before starting cleaning or disinfection work. Do not allow any liquids to penetrate the inside of the housing. Do not spray clean/spray disinfect the housing. Only plug the device back in if it is completely dry, both inside and outside.
Maintenance Eppendorf BioSpectrometer® basic English (EN) 8.1.1 Cleaning the cuvette shaft cover If you would like not only to clean the directly accessible surface of the cuvette shaft cover, you can remove the cover. Do not soak the cuvette shaft cover in cleaning agent. Clean the cuvette shaft cover as described. 1. Lift the cuvette shaft cover with one hand. 2.
Maintenance Eppendorf BioSpectrometer® basic English (EN) 8.2 Disinfection/Decontamination DANGER! Electric shock as a result of penetration of liquid. Switch off the device and disconnect the power plug before starting cleaning or disinfection work. Do not allow any liquids to penetrate the inside of the housing. Do not spray clean/spray disinfect the housing. Only plug the device back in if it is completely dry, both inside and outside. 1.
Maintenance Eppendorf BioSpectrometer® basic English (EN) Abb. 8-1: Inside of the filter box lid (sample) Fig.
Maintenance Eppendorf BioSpectrometer® basic English (EN) 8.3.1.1 Checking photometric accuracy 1. Select the Spectrometer unit function in the Device calibration group and confirm your selection with enter. 2. Select whether you want to check the wavelength systematic error or photometric accuracy, and confirm with enter. Press [Next >] to switch to the measurement. 3. Follow the instructions on the device display and start by measuring the A0 blank filter, and then the first test filter A1.
Maintenance Eppendorf BioSpectrometer® basic English (EN) 4. Results display after measuring a test filter for testing photometric accuracy. Measure the other two test filters A2 and A3. 5. Results display after measuring all 3 test filters for testing the photometric accuracy. With the and keys, you can view the results for the different test filters again. Press [Finish] to complete the test. 6. Compare the average values and CV values to the supplied table.
Maintenance Eppendorf BioSpectrometer® basic English (EN) 8.3.2 Device self-test The frequency of the automatic self-test (approx. 1 minute) can be set using the Device settings function (see Device settings on p. 64). The factory setting for Self-test interval is "Weekly".
Maintenance Eppendorf BioSpectrometer® basic English (EN) 8.5 Decontamination before shipment If you are shipping the device to the authorized Technical Service for repairs or to your authorized dealer for disposal please note the following: WARNING! Risk to health from contaminated device 1. Follow the instructions in the decontamination certificate. You find it as a PDF file on our website (www.eppendorf.com/decontamination). 2. Decontaminate all the parts you would like to dispatch. 3.
Troubleshooting Eppendorf BioSpectrometer® basic English (EN) 9 Troubleshooting 9.1 General errors Error Measuring results are imprecise. Possible cause Remedy • Reagent is past its shelf life. Ensure that the reagent is still within its shelf life and properly prepared. • Reagent has not been prepared properly. Use clean demineralized water of adequate quality • Pipetting is not correct. Ensure that the pipette is calibrated and that • Incubation procedure before measurement is incorrect.
Troubleshooting Eppendorf BioSpectrometer® basic English (EN) 9.2 Error messages You can exit device displays with error messages using the [OK] softkey. System errors require an evaluation by the Technical Service. These errors are shown in English (System error …). Please contact Technical Service in these cases. Other error messages, for which you can carry out troubleshooting measures, are illustrated in the table below. Problem Self test failed.
Troubleshooting Eppendorf BioSpectrometer® basic English (EN) Problem The following parameter values are not defined in General Method Parameter: Cause Solution Select a different parameter from the • When opening a method with existing list. If necessary, program a parameters which access General new list entry in General Method Method Parameter, the system Parameter in order to be able to use it determined that at least one when programming a method.
Troubleshooting Eppendorf BioSpectrometer® basic English (EN) Problem Cause Solution Calculation not possible because of division by zero. Absorbance result or Formula "b" parameter is zero. • An absorbance result was divided by a "zero" value during the evaluation of a Division type method (Dual wavelength method group). This is not mathematically permissible. Check the reagents and samples used There is only one measurement left to be performed in this series of measurement.
Troubleshooting Eppendorf BioSpectrometer® basic English (EN) 9.3 Result flags Warnings and error messages for results are displayed in the bottom right of the help box. The header bar of the Help box is highlighted yellow for warnings and red for error messages. Warnings: Decide whether the result is useful for you while taking the displayed warning into consideration. Error messages: No result is displayed; the reason is shown in the error message.
Troubleshooting Eppendorf BioSpectrometer® basic English (EN) Problem Cause Solution The coefficient of determination for the regression evaluation of the standard series is < 0.8. Use the sample results with the • For methods with evaluation of reservation mentioned or repeat the standard series via the regression measurement of the standard series procedure: If the regression and samples.
Troubleshooting Eppendorf BioSpectrometer® basic English (EN) Problem • Calculation not possible because of division by zero. Absorbance result is zero. • Calculation error. Division by zero. Calculation not possible because of division by zero. Absorbance result or parameter formula b is zero. Cause Solution Check the reagents and samples used • The evaluation required dividing by and repeat the measurement. an absorbance result with the value of "zero". This is not mathematically permissible.
Troubleshooting Eppendorf BioSpectrometer® basic English (EN)
Transport, storage and disposal Eppendorf BioSpectrometer® basic English (EN) 10 Transport, storage and disposal 10.1 Transport Use the original packaging for transport.
Transport, storage and disposal Eppendorf BioSpectrometer® basic English (EN)
Technical data Eppendorf BioSpectrometer® basic English (EN) 11 Technical data 11.1 Power supply Power supply 100 V to 240 V ±10 %, 50 Hz to 60 Hz Overvoltage category II Degree of pollution 2 Power consumption Maximum power consumption according to name plate: 25 W Approx. 15 W during operation Approx. 5 W with the display dimmed Permitted mains interruption Approx. 10 ms at 90 V Approx. 20 ms at 230 V Protection class I Fuses T 2.5 A/250 V, 5 mm × 20 mm (2 pcs.) 11.
Technical data Eppendorf BioSpectrometer® basic English (EN) 11.
Technical data Eppendorf BioSpectrometer® basic English (EN) 11.
Technical data Eppendorf BioSpectrometer® basic English (EN)
Evaluation procedure Eppendorf BioSpectrometer® basic English (EN) 12 Evaluation procedure This chapter describes the evaluation procedures available in the method programs as well as the calculation of a dilution using the device software. When comparing the measuring results to the results of other photometers/ spectrophotometers, note that the values may be dependent on the bandwidth of the devices.
Evaluation procedure Eppendorf BioSpectrometer® basic English (EN) 12.1.3 Cuvette correction All absorbance values which are used for result calculation are standardized to the cuvette layer thickness of 10 mm. If a cuvette with a different path length is used, this path length must be defined in the cuvette parameter. In this case, the measured absorbances are corrected to match measuring results with a cuvette layer thickness of 10 mm before converting them to sample results.
Evaluation procedure Eppendorf BioSpectrometer® basic English (EN) 12.2 C Evaluation with factor or standard Au F C = calculated concentration. A = absorbance. F = factor. The factor is programmed in the parameter list and can be modified. It always relates to an optical path length of the cuvette of 10 mm. If you change the Cuvette parameter the device will take the modification into account when calculating the results. Therefore you do not need to change the factor for the evaluation.
Evaluation procedure Eppendorf BioSpectrometer® basic English (EN) • Use the "linear regression" procedure for calibration lines. • With curvilinear gradients, test which evaluation procedure (quadratic regression, cubic regression, spline interpolation) produces the function that is most suitable to the standard evaluation.
Evaluation procedure Eppendorf BioSpectrometer® basic English (EN) 12.5 Special evaluation procedures for nucleic acids and protein UV This section covers the evaluation of nucleic acids or proteins in the Nucleic acids and Proteins direct UV method groups, as well as the corresponding biomolecular components in the Dye labels method group. 12.5.
Evaluation procedure Eppendorf BioSpectrometer® basic English (EN) 12.5.3 Conversion to molar concentrations and nucleic acid quantities The conversion only can be applied to nucleic acids and dye methods with nucleic acids as biomolecule components. It is realized in the process results/More calculations method step. 12.5.3.1 Calculation of amount Application: calculating the amount (mass) of nucleic acid in the total sample volume.
Evaluation procedure Eppendorf BioSpectrometer® basic English (EN) • For dsDNA the calculation of the molar concentration is based on the assumption of a double-stranded nucleic acid. For the ssDNA, RNA and Oligo methods, a single-stranded nucleic acid is assumed. • For methods which have been reprogrammed via in the Routine main group, Nucleic acids method group, always double-stranded nucleic acids are assumed for calculating the molar concentration. 12.5.
Evaluation procedure Eppendorf BioSpectrometer® basic English (EN) 12.6.2 Calculation of the FOI As a value for the ratio of dye molecules to the number of nucleotides in the nucleic acid the frequency of incorporation (FOI) is calculated and displayed for the dye methods. The calculation can be selected for two different result units: MOLECULE dye/kb unit AYYY 106 u MM nt u H Dye AXXX u FNA FOI pmole/μg DNA (or RNA) unit AYYY FOI H Dye u 109 AXXX u FNA AYYY = absorbance of the dye.
Evaluation procedure Eppendorf BioSpectrometer® basic English (EN) 12.7 Dual wavelength For methods of the Dual Wavelength group absorbances that were measured at two wavelengths can be calculated with each other before the calculated absorbance is evaluated further with the factor or standard. To determine the calculated absorbance a division or subtraction evaluation can be defined in the parameters: Acalc a u A1 uc d b u A2 Acalc > a u A1 b u A2 @u c d A1, A2 = measured absorbance.
Evaluation procedure Eppendorf BioSpectrometer® basic English (EN)
Ordering information Eppendorf BioSpectrometer® basic English (EN) 13 Ordering information Order no. (International) Order no. (North America) 6135 000.009 – 6135 000.017 6135000017 6135 928.001 6135928001 6138 000.018 6138000018 6135 011.000 6135 010.004 6135 012.007 6135010004 0013 021.566 952010409 0030 106.300 952010051 0030 106.318 952010069 0030 079.345 0030079345 0030 079.353 0030079353 4308 078.
Ordering information Eppendorf BioSpectrometer® basic English (EN)
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