oSpectrometer® nual gEN) manual kinetic 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® kinetic English (EN) Table of contents 1 Operating instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.1 Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.2 Danger symbols and danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Table of contents Eppendorf BioSpectrometer® kinetic English (EN) 6 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.1 Selecting a method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2 Photometry method description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2.
Table of contents Eppendorf BioSpectrometer® kinetic English (EN) 11 Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.1 Power supply. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.2 Ambient conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.
Table of contents Eppendorf BioSpectrometer® kinetic English (EN)
Operating instructions Eppendorf BioSpectrometer® kinetic English (EN) 1 1.1 Operating instructions 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® kinetic English (EN) Depiction Meaning Press this softkey to perform the described action. or [Copy] Additional information 1.
Product description Eppendorf BioSpectrometer® kinetic English (EN) 2 2.1 Product description 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® kinetic English (EN) 2.3 Features The BioSpectrometer kinetic 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® kinetic English (EN) 3 3.1 Safety Intended use The BioSpectrometer kinetic is to be used in molecular biology, biochemistry and cell biology research laboratories. The BioSpectrometer kinetic is exclusively intended for use indoors. All country-specific safety requirements for operating electrical equipment in the laboratory must be observed.
Safety Eppendorf BioSpectrometer® kinetic 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® kinetic English (EN) 3.3.2 Damage to device NOTICE! Damage from the use of aggressive chemicals. Do not use any aggressive chemicals on the device or its accessories, such as strong and weak bases, strong acids, acetone, formaldehyde, halogenated hydrocarbons or phenol. If the device has been contaminated by aggressive chemicals, immediately clean it by means of a mild cleaning agent. NOTICE! Damage to the device from fumigating with aggressive chemicals.
Safety Eppendorf BioSpectrometer® kinetic English (EN) NOTICE! Material damage from incorrect use. Only use the product for its intended purpose as described in the operating manual. Ensure adequate material resistance when using chemical substances. In case of doubt, contact the manufacturer of this product. NOTICE! Damage as a result of incorrect packing. Eppendorf AG is not liable for damage caused by improper packing.
Installation Eppendorf BioSpectrometer® kinetic English (EN) 4 4.1 Installation 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. 9). Check all parts for any transport damage. 4.
Installation Eppendorf BioSpectrometer® kinetic 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. 9). 2. Connect the printer cable to the printer and tighten the locking screws as well. 3.
Installation Eppendorf BioSpectrometer® kinetic English (EN) DIP SW-3 Meaning 5 (OFF) Baud 6 (ON) Rate 7 (ON) Select 8 (ON) = 9600 bps 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. 9). 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® kinetic English (EN)
Operation Eppendorf BioSpectrometer® kinetic English (EN) 5 5.1 Operation Overview of operating controls Abb. 5-1: Control panel of the BioSpectrometer kinetic Fig. 5-1: Key: Control panel of the BioSpectrometer kinetic 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.
Operation Eppendorf BioSpectrometer® kinetic English (EN) Key: Function Move the cursor to the left, right, up, down. • Navigation between input fields. • and keys inside an entry field: Navigate within the character string. • and keys in a result display: Navigate between the sample results of the series of measurement. • and keys within a graph: Navigate on the x-axis of the graph, e.g. for displaying the wavelength-dependent absorbance values in a scan.
Operation Eppendorf BioSpectrometer® kinetic 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® kinetic English (EN) Eppendorf Hellma ® TrayCell * UVette ® µCuvette G1.0 Cuvettes Ultra-micro Semi-micro 70 µL 400 µL Macro Basic area 12.5 mm × 12.5 mm Min. overall height 36 mm Min. filling level Light path Max. height of base 10 mm 8.5 mm 7 mm 0 mm Min.
Operation Eppendorf BioSpectrometer® kinetic English (EN) 5.3.2 Measuring procedure 5.3.2.1 Selecting a method Use the cursor keys to select the desired method and call up the method with the enter key. For an overview and a detailed description of the methods, refer to the next chapter (see Methods on p. 29). Wizard: The wizard at the top of the display will take you through the method procedure step-by-step.
Operation Eppendorf BioSpectrometer® kinetic English (EN) 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). The display always marks the standard that is to be measured next. Use the [Graph] resp. [Table] softkey to change the result view. Press [Next] to accept the evaluation calculated from the standard results. 5.3.2.
Operation Eppendorf BioSpectrometer® kinetic English (EN) 5.3.2.5 Finalizing the method 1. Press [Finish], to complete the measuring series and return to the method selection. 2. After all measurements have been completed, switch off the device and close the cuvette shaft cover to protect the cuvette shaft from contamination. 5.3.2.6 Optional: process results For some methods, you can postprocess the results in the process results method step.
Operation Eppendorf BioSpectrometer® kinetic 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® kinetic English (EN) Measured values typical for Eppendorf for temperature control in closed cuvettes, with closed cuvette shaft covers, are shown in the following tables. The temperature was measured in the measuring solution; the ambient temperature was 24.5°C. Tab.
Operation Eppendorf BioSpectrometer® kinetic English (EN) • For efficient temperature control, the volume of the measuring solution in the cuvette should not project beyond the edge of the cuvette holder. • To speed up the measuring procedure during series measurements, you can pre-cool cuvettes with reagents in a thermostat outside the BioSpectrometer before inserting the cuvette in the cuvette holder and adding the sample.
Methods Eppendorf BioSpectrometer® kinetic English (EN) 6 6.1 Methods 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® kinetic English (EN) You can create new methods in all folders using . In Favorites, you can create your own folders (e.g., to allocate folders to specific people), and rename and delete the folders. 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.
Methods Eppendorf BioSpectrometer® kinetic 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® kinetic English (EN) • Correction of the influence of the dye spectrum on the accuracy of the biomolecule measurement is possible. • Partial turbidity correction can be performed via the Background parameter. • Additional information on the purity of the measured materials: Ratios A260/A280 and ratios A260/A230 (ratio values only for nucleic acids), absorbance wavelength spectrum.
Methods Eppendorf BioSpectrometer® kinetic 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.
Methods Eppendorf BioSpectrometer® kinetic English (EN) Parameter Entry Explanation Unit Unit for the concentration result. Selection: In the preprogrammed methods of the Routine group, the mg/mL | μg/mL | ng/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® kinetic 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: Simple kinetics, Advanced kinetics, Dual wavelength, Factor. For the Dye labels method group the factors are not entered into the method procedure individually.
Methods Eppendorf BioSpectrometer® kinetic 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® kinetic English (EN) Parameter Entry Explanation 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).
Methods Eppendorf BioSpectrometer® kinetic English (EN) Parameter Entry Explanation Measuring procedure Selection: lin.regr. | endpoint | two point For kinetic methods only. "Linear regression": Measurement over several periods in set time intervals within a defined period of time. Evaluation via linear regression of the absorbance time graph within the measuring time. Absorbance result: ΔA/min. "Endpoint": Measurement of a measuring point after a defined period of time. Absorbance result: A.
Methods Eppendorf BioSpectrometer® kinetic 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® kinetic 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® kinetic 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® kinetic 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® kinetic 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® kinetic English (EN) Result image with dilution and ID Result image with dilution and sample ID
Methods Eppendorf BioSpectrometer® kinetic English (EN) 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® kinetic English (EN) Method group Results display Explanation Multi λ Results display: • Absorbance values at the wavelengths Additional data ([Data] softkey): • Only for dilution or with cuvettes other than 10 mm: absorbance value before the conversion. Scan Results display: • Scan (graph with absorbance wavelength display) • Navigate between the measuring points in the graph with and .
Methods Eppendorf BioSpectrometer® kinetic English (EN) Method group Results display Explanation Proteins direct UV 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® kinetic English (EN) Method group Results display Explanation Bacterial density Results display: • Calculated result with absorbance at the measuring wavelength. • If activated in the parameters: Scan. Navigate between the measuring points in the graph with and . Basic main group Factor, standard Analog to Protein direct UV (see above) Results display: • Concentration result with absorbance at the measuring wavelength.
Methods Eppendorf BioSpectrometer® kinetic 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.
Methods Eppendorf BioSpectrometer® kinetic 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® kinetic 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]. 6.4.6 Process results: Options Zoom Press the [Zoom] softkey and select one of the following versions.
Methods Eppendorf BioSpectrometer® kinetic English (EN) Variant [spectra-0]: Same as the [spectra] variant, with one exception: The lower limit of the displayed section of the y-axis always equals "0 A". Variant [free]: User-defined values for interval limits can be entered for both axes. Navigation between the entry fields by means of the cursor keys ( , , ). For all 3 versions, the [reset zoom] softkey will bring you back to the original display of the spectrum.
Methods Eppendorf BioSpectrometer® kinetic 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® kinetic English (EN) Examples: λ grid: 100 nm, min. Δ Abs: 0.050: The peak is not detected because the λ grid is too large: The absorbance values on the left edge of the grid are higher than the absorbance of the peak. λ grid: 20 nm, min. Δ Abs: 0.200: The peak is not detected because the predetermined value for min. Δ abs is too high. The difference of the absorbance of the peak and the lowest absorbance in the grid is less than 0.2 A. λ grid: 20 nm, min. Δ Abs: 0.
Methods Eppendorf BioSpectrometer® kinetic English (EN) Linear regression For kinetic methods evaluated using the "Linear regression" measurement method, you can subsequently define the start and end times for the regression evaluation in the process results method step. Selecting measuring points • [First pt.] softkey: Define the first measuring point for the regression evaluation. Select the measuring point using and . Confirm with enter. • [Last pt.
Methods Eppendorf BioSpectrometer® kinetic English (EN) Data export The data will be transferred as Excel files (.xls) and can be read with Excel versions Excel 97 and later. For each of the selected data packets, a worksheet is created in Excel. The file name consists of the method name, the time and the date of the measuring series. Select export version If no USB stick is connected, the first variant cannot be selected. Export to USB stick 1.
Methods Eppendorf BioSpectrometer® kinetic English (EN) 6.4.8 Finish the series of measurements After the print & export method step has been finished, you can start a new series of measurements using the selected method or select a new method. Finish the series of measurements and start a new series of measurements • [Next >] softkey: Call up new series method step • [New] softkey: Call up measure samples method step and start a new series of measurements.
Methods Eppendorf BioSpectrometer® kinetic English (EN)
Functions Eppendorf BioSpectrometer® kinetic English (EN) 7 7.1 Functions 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® kinetic 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® kinetic English (EN) 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. In this example, parameter groups are summarized for various dyes (dye components for the dye methods) and stored under a name. With this name, the required parameter group can be imported into the method program during the editing of a dye method. Display: • Left: Name of the dye.
Functions Eppendorf BioSpectrometer® kinetic English (EN) When programming a method of the Dye labels or Proteins direct UV method groups, you can access the entries in General Method Parameter: Select the name of the dye to import the corresponding parameter group into the method program. By using the "edit" selection of the "Nucleic acid" parameter, you also can get directly to the General Method Parameter function and view and edit the parameters. Tab.
Functions Eppendorf BioSpectrometer® kinetic English (EN) • Specifications for proteins which are not preset at the factory can be determined in the expasy database: http://www.expasy.org/tools/protparam.html. • A table with A1% values for many proteins can also be found in: C.N.Pace et al., Protein Science (1995), 4: 2411–2423 (Table 5). The A1% values must be multiplied by 0.1 to return the required A0.1% values. 7.1.
Functions Eppendorf BioSpectrometer® kinetic English (EN) 7.1.5 Device calibration Information on checking the device is provided separately (see Checking the device on p. 66). 7.1.6 Info The Copyright menu item contains license information on the Open Source software.
Maintenance Eppendorf BioSpectrometer® kinetic English (EN) 8 8.1 Maintenance 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® kinetic English (EN) 3. Clean the cover with a cloth or lint-free cotton swab dampened with a mild cleaning agent. 4. Slide the locking pin back into the housing as far as it will go. The locking pin has completely disappeared in the housing. When the photometer is not being used, close the cuvette shaft using the blue cuvette shaft cover to protect it from dust and other contamination. 8.
Maintenance Eppendorf BioSpectrometer® kinetic English (EN) 8.3.1 Checking the spectrometer unit Eppendorf offers a filter kit (BioSpectrometer reference filter kit) for checking the photometric accuracy and wavelength systematic error. The kit contains one blank filter (A0) and three filters (A1, A2 and A3) for checking the photometric accuracy, and 3 filters for checking the wavelength systematic error in the range of 260 nm to 800 nm. The filter absorbances are measured against blank filter A0.
Maintenance Eppendorf BioSpectrometer® kinetic English (EN) Abb. 8-1: Inside of the filter box lid (sample) Fig.
Maintenance Eppendorf BioSpectrometer® kinetic English (EN) 8.3.1.1 Checking photometric accuracy 1. Select the Spectrometer unit function in the Device calibration group and confirm 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® kinetic 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® kinetic English (EN) 8.3.2 Checking the thermal module • Carry out an inspection at approx. 20 °C. • Make sure that the cuvette shaft is empty for the check. 1. Select the Temperature unit function in the Device calibration group and confirm with enter. 2. Make sure the cuvette holder is empty, and start the test procedure with [Next >]. The following temperature control test at 5 temperatures takes approx. 30 minutes.
Maintenance Eppendorf BioSpectrometer® kinetic English (EN) 8.3.3 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. 63). The factory setting for Self-test interval is "Weekly".
Maintenance Eppendorf BioSpectrometer® kinetic 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.
Maintenance Eppendorf BioSpectrometer® kinetic English (EN)
Troubleshooting Eppendorf BioSpectrometer® kinetic English (EN) 9 9.1 Troubleshooting 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 for preparation if required. • Pipetting is not correct.
Troubleshooting Eppendorf BioSpectrometer® kinetic English (EN) Error Possible cause The measuring results are not correct. • The method has not been programmed correctly. Remedy Ensure that the method parameters are entered correctly. • The standard solution has Ensure that the correct standard is used not been prepared and that the measuring solution for the correctly. standard is prepared correctly. 9.2 • The absorbance of the reagent is drifting.
Troubleshooting Eppendorf BioSpectrometer® kinetic English (EN) Problem Cause Solution Blank measurement: An intensity on a pixel that influences the main, auxiliary or scan wavelength is too low. • The absorbance of the blank solution Check the blank solution and used for the blank measurement is too remeasure the blank if required. high. For scans: Match the wavelength • Incorrect or turbid blank solution. range to the sample spectrum.
Troubleshooting Eppendorf BioSpectrometer® kinetic English (EN) Problem Cause Solution At least two of the entered standard concentrations are identical. Correct the standard concentrations. • See the error text. Enter the standard concentrations so that the first standard receives the lowest concentration and the other standard concentrations form an increasing sequence.
Troubleshooting Eppendorf BioSpectrometer® kinetic English (EN) Problem Cause Solution Invalid zoom interval! Error in the process results method step Please observe the stated limits in the in the Zoom mode. zoom procedure. Permissible zoom range for the wavelength scale: • Wavelength interval at least 10 nm • Entries for wavelengths only within the range programmed in the parameters for the method. Permissible zoom range for the absorbance scale: • Absorbance interval at least 0.
Troubleshooting Eppendorf BioSpectrometer® kinetic 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® kinetic English (EN) Problem Cause Solution The coefficient of determination for the regression evaluation of the standard series is < 0.8. • For methods with evaluation of standard Use the sample results with the series via the regression procedure: If the reservation mentioned or repeat regression evaluation for the standard the measurement of the standard series was nonlinear, but the standard series and samples.
Troubleshooting Eppendorf BioSpectrometer® kinetic English (EN) Problem Cause Solution Measure the sample at ambient temperature within the specified range (15°C to 35°C). If the warning nevertheless appears, contact Eppendorf Service. During the kinetics measurement, the temperature was outside of the allowable range. • Ambient temperature outside the specified range. • The temperature control is faulty. Absorbance at the measuring wavelength is too high.
Transport, storage and disposal Eppendorf BioSpectrometer® kinetic English (EN) 10 10.1 Transport, storage and disposal Transport Use the original packaging for transport.
Transport, storage and disposal Eppendorf BioSpectrometer® kinetic English (EN)
Technical data Eppendorf BioSpectrometer® kinetic English (EN) 11 11.1 Technical data 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: 50 W Approx. 30 W during operation Approx. 5 W with the display dimmed and temperature control switched off Permitted mains interruption Approx. 10 ms at 90 V Approx. 20 ms at 230 V Protection class I Fuses T 2.
Technical data Eppendorf BioSpectrometer® kinetic English (EN) 11.
Technical data Eppendorf BioSpectrometer® kinetic English (EN) 11.6 Further technical parameters Cuvette material For measurements in the UV: Quartz glass or UV transparent plastic (Eppendorf UVette, 220 nm to 1600 nm) For measurements in the visible range: Glass or plastic material Cuvette shaft 12.5 mm × 12.5 mm, tempered Overall cuvette height Min. 36 mm Height of the light beam in the cuvette 8.
Technical data Eppendorf BioSpectrometer® kinetic English (EN) 11.
Evaluation procedure Eppendorf BioSpectrometer® kinetic 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® kinetic 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® kinetic English (EN) The factor is either entered directly as a parameter during the "Factor" evaluation procedure or calculated during the "Standard" evaluation procedure (evaluation with a standard concentration): F CS AS F = calculated factor. CS = concentration of the standard (enter as parameter). AS = measured absorbance of the standard.
Evaluation procedure Eppendorf BioSpectrometer® kinetic 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® kinetic 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® kinetic 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® kinetic 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® kinetic English (EN) 12.6 Special evaluation procedures for the dye methods 12.6.1 Calculating the factor for the dye from the absorbance coefficient For the dye methods the concentration of the dye is calculated using a factor from the measured absorbance (see Evaluation with factor or standard on p. 90). The factor is entered for each dye in the General Method Parameter/Dyes function. Alternatively, you can enter the absorbance coefficient.
Evaluation procedure Eppendorf BioSpectrometer® kinetic English (EN) 12.6.3 Conversion to amounts of dye The quantity (dimensions) of dye in the entire sample volume is calculated in the process results/More calculations method step. M C u VP ,total M = calculated total amount (mass) of dye in the sample tube. Unit: pmol. C = dye concentration calculated from the measurement. Unit: pmol/μL. VS, total = total volume of the sample in the sample tube; entered by the user under More calculations.
Evaluation procedure Eppendorf BioSpectrometer® kinetic English (EN) 12.8 Kinetics An absorbance value A or an absorbance difference standardized to a minute ΔA/min is determined using the measurement procedures available for selection. This determined absorbance value is entered in the concentration calculation using a factor or (only for Advanced kinetics) with an end-point standard. 12.8.
Evaluation procedure Eppendorf BioSpectrometer® kinetic English (EN) If the time frame for the kinetic evaluation with linear regression is reduced in the process results method step, the reduced measuring window is automatically used for calculating the reagent blank which was incorporated in the calculation of this sample result. That means that the reagent blank is recalculated (only) for the calculation of this sample result.
Evaluation procedure Eppendorf BioSpectrometer® kinetic English (EN)
Ordering information Eppendorf BioSpectrometer® kinetic English (EN) 13 Ordering information Order no. (International) Order no. (North America) 6135 000.009 – 6135 000.017 6135000017 6136 000.002 – 6136 000.010 6136000010 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® kinetic English (EN)
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