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Содержание NanoQuant Plate

Страница 1: ...NanoQuant Plate Quick Guide ...

Страница 2: ... Requirements 5 1 5 Applications 6 2 Measurement Procedure 7 2 1 Software Installation Procedure 7 2 2 Performing a Measurement 7 2 3 File menu 15 2 4 Quality Control of NanoQuant Plate 17 3 NanoQuant Plate 19 3 1 Parameters 19 3 2 Handling and Cleaning 21 3 3 Applying Samples 22 3 4 NanoQuant Plate Disinfection 23 4 Calculations 25 4 1 Calculation of Nucleic Acid Concentration 25 5 About the Quic...

Страница 3: ...tomatically and the results are displayed in an Excel sheet A blanking measurement including an integrated reference wavelength at the beginning of the measurement procedure functions simultaneously as a quality control check for the entire plate and indicates any pipetting or cleaning mistakes The plate has been designed to meet the requirements of research laboratories working with various types...

Страница 4: ...pace Requirements 700 MB 1 GB Monitor Super VGA Graphics Resolution 1024 x 768 1280 x 1024 Color Depth 256 Mouse Microsoft mouse or compatible pointing device Communication 1 x USB 2 0 2 x USB 2 0 1 x RS232 Serial Devices 1 x CD ROM drive Windows Vista DirectX 9 graphics and 32 MB of graphics memory for Home Basic 128 MB of graphics memory plus WDDM support for all other versions Windows 7 DirectX...

Страница 5: ... PRO the filter positions must remain in the order in which they were delivered The original filter positions guarantee the fastest filter switching for well wise measurements The Infinite M200 PRO can be used immediately for measurement without any calibration of the monochromator Infinite M1000 Infinite F500 An Infinite M1000 reader REF 30061442 with main firmware version V2 0 or higher An Infin...

Страница 6: ...eptable If this ratio is lower than 1 8 it may indicate the presence of proteins or other contaminants If this is the case an additional purification step procedure is recommended Labeling Efficiency Infinite M200 PRO and Infinite M1000 only Working with nucleic acids labeled with fluorescent dyes requires samples of high quality Besides common nucleic acid quantification and nucleic acid purity c...

Страница 7: ...ise Installation Wizard 5 When installation is successful exit the Installation window 6 Switch on and connect to Infinite instrument Note i control is delivered with the Infinite reader series 2 2 Performing a Measurement For applications using the NanoQuant Plate a tab called Applications is implemented in i control software so that all measurements can be performed quickly and easily 1 Start i ...

Страница 8: ...g on the connected instrument the wavelengths used for measurement are selected automatically make sure that the correct filters are properly installed and defined on the filter slides of the Infinite F200 PRO and Infinite F500 instrument Instrument Bandwidth at 260 nm Bandwidth at 280 nm Infinite M200 PRO 5 nm 5 nm Infinite F200 PRO 5 nm 3 nm Infinite M1000 5 nm 5 nm Infinite F500 5 nm 3 nm 8 Sel...

Страница 9: ...ue Labeling Efficiency Blanking results date and time samples positions that were selected for blanking blanking range and maximum CV are displayed next to the plate preview in the measurement stripe and saved until the instrument is disconnected 13 When the blanking measurement has been completed successfully the plate is moved out automatically The plate is now ready for sample application and a...

Страница 10: ...values of each sample at all relevant wavelengths are also displayed Figure 3 Overview of Excel result sheet 17 Once the measurement procedure is finished the plate is moved out automatically A pop up message appears asking if the user wishes to perform another measurement If additional identical measurements are to be performed wipe oﬀ any sample residues from the previous measurement and apply n...

Страница 11: ...clean the NanoQuant Plate thoroughly and store it appropriately 19 Once i control has been closed the Infinite reader can be switched off Note Excel workbooks containing the measurement results are not saved automatically This has to be done by the user It is possible to save the method script created in i control for further use by clicking Save in the File menu ...

Страница 12: ...the correct fluorophores are selected the samples can be applied into the NanoQuant Plate Individual Blanking and Average Blanking The user may choose between two distinct blanking options Average Blanking and Individual Blanking set by default For Average Blanking choose which wells are to be used for blanking by dragging a frame around the respective sample positions in the plate preview It is g...

Страница 13: ...single blanking value of the same well instead of one average blank Figure 6 Select Individual Blanking and click the Start Blanking button to start individual blanking It is recommended to use Individual Blanking as the standard blanking option in order to obtain the most precise and reliable results Figure 7 Individual Blanking information in the Excel result sheet Note The CV value displayed in...

Страница 14: ...sired sample names By clicking the Clear button all inserted Sample IDs are deleted Sample IDs are displayed in the summary sheet Click Save to store the entries and click Close to complete the procedure Sample IDs may also be copied from Excel sheets and pasted into sample ID list Show Raw Data To view all measurement results as raw values the NanoQuant tab in the Results Presentation dialog box ...

Страница 15: ...es and the automatically calculated sample concentrations and ratio values Figure 8 Raw Data Output in the Excel results summary 1 Nucleic acid quantification 2 Labeling efficiency 2 3 File menu i control Toolbar Start a measurement Connect or disconnect an instrument Move plate out Move plate in Show or hide the info pane Open the i control help file Sample The following sample types can be selec...

Страница 16: ... from the drop down menu Figure 9 Selecting and adding new samples Note For the Infinite F200 PRO and Infinite F500 the correct filters must always be used and defined correctly in the filter slide If a wavelength is selected that is not available because the filter is not in the slide an error message appears Figure 10 Editing or creating new markers Note The selected sample in Edit Sample must a...

Страница 17: ...ells are highlighted in pink and an error message appears requesting the user to repeat blanking measurement In this case single wells are displayed in pink indicating these wells as diverging after blanking These wells differ from the calculated CV by 10 meaning that these quartz wells are dirty due to lint fingerprints etc Figure 11 Color code for blanking pink wells are out of range ...

Страница 18: ...elow by dragging a frame around them The newly selected wells will appear white the diverging wells change from pink to purple and all other wells turn blue to indicate that they are unused Figure 12 Color code for blanking procedure when re selected purple wells indicate formerly pink wells 2 Move the plate out repeat the cleaning procedure and apply fresh blanking samples into the plate and star...

Страница 19: ...eight 14 6 mm Plate length 127 8 mm Weight 160 g Wavelength settings Infinite M200 PRO 260 nm 5 nm bandwidth 280 nm 5 nm bandwidth 310 nm 5 nm bandwidth reference Infinite M1000 260 nm 5 nm bandwidth 280 nm 5 nm bandwidth 310 nm 5 nm bandwidth reference Infinite F200 PRO Infinite F500 260 nm 5 nm bandwidth 280 nm 3 nm bandwidth 340 nm 10 nm bandwidth reference Measurement time of whole plate Quant...

Страница 20: ... for λ 315 nm 0 3 nm for λ 315 nm Absorbance bandwidth Infinite M200 PRO 5 nm for λ 315 nm 9 nm for λ 315 nm Absorbance bandwidth Infinite F200 PRO Depending on filters used Measurement absorbance Detection limit DNA concentration 1 ng μl dsDNA Reproducibility of one sample 50 μg ml 1 CV Ratio 260 280 50 μg ml 0 07 Precision 260 nm 0 2 Accuracy 260 nm 0 5 Figure 13 Technical drawing of the NanoQua...

Страница 21: ...ethanol and clean the inner and outer surfaces of the NanoQuant Plate 2 Moisten a piece of cotton or Kimwipe with distilled water and clean both sides of each quartz lens on the NanoQuant Plate 3 Wipe off any excess liquid with a dry Kimwipe After cleaning store the plate in a dirt free and lint free place No lint nor any kind of dirt or streaks should be on the quartz lenses Any contamination can...

Страница 22: ...istent application of the samples Figure 14 Applying samples onto the NanoQuant Plate If you need help applying the samples precisely into the wells with a multi channel pipette use our pipetting aid Place the pipetting aid with the indentation downward onto the quartz wells The pipetting aid must be secured on the steel pins Figure 15 NanoQuant Plate with pipetting aid Apply the samples into the ...

Страница 23: ...ngth and therefore to imprecise OD values 3 4 NanoQuant Plate Disinfection All parts of the NanoQuant Plate that come in contact with patient samples positive control samples or hazardous material must be treated as potentially infectious areas Spray or apply 70 ethanol which is commonly used for laboratory cleaning over the whole plate After an exposure time of 5 minutes dry the NanoQuant Plate w...

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Страница 25: ...tration according to the Lambert Beer law including the reference values A ε d c A Absorbance ε Molarity Extinction Coefficient L mol 1 cm 1 d Distance path length in cm c Concentration mol L 1 Calculation of DNA concentration DNA purity is automatically performed in the background as described in the following Generally in analytical chemistry the absorbance A is defined as Aλ log10 𝑰𝟎 𝑰 OD where...

Страница 26: ...nite F500 of all wells used for blanking is subtracted from the average absorbance value at 260 nm or 280 nm The relative variation of the wells used for average blanking must be below 10 in order to be able to start a measurement Infinite M200 PRO and Infinite M1000 Abs blank average Abs 260 average Abs 310 average OD Infinite F200 PRO and Infinite F500 Abs blank average Abs 260 average Abs 340 a...

Страница 27: ...alculations based on Average Blanking The well specific absorbance value at 310 nm for Infinite M200 PRO and Infinite M1000 340 nm for Infinite F200 PRO and Infinite F500 is subtracted from the corresponding absorbance value at 260 nm Each well used for sample measurement is then blanked with the average blanking value Infinite M200 PRO Infinite M1000 Abs A1 Abs 260 A1 Abs 310 A1 Abs blank average...

Страница 28: ...PRO Infinite M1000 Abs A1 Abs 260 A1 Abs 310 A1 Abs blank average OD Abs A2 Abs 260 A2 Abs 310 A2 Abs blank average OD Abs B1 Abs 260 B1 Abs 310 B1 Abs blank average OD etc Infinite F200 PRO Infinite F500 Abs A1 Abs 260 A1 Abs 340 A1 Abs blank average OD Abs A2 Abs 260 A2 Abs 340 A2 Abs blank average OD Abs B1 Abs 260 B1 Abs 340 B1 Abs blank average OD etc The absorbance values at 280 nm are also ...

Страница 29: ...n the NanoQuant Plate Note Every effort has been made to avoid errors in text and diagrams however Tecan Austria GmbH assumes no responsibility for any errors that may appear in this publication It is the policy of Tecan Austria GmbH to improve products as new techniques and components become available Tecan Austria GmbH therefore reserves the right to change specifications at any time with approp...

Страница 30: ...emarks of Tecan Group Ltd Männedorf Switzerland Kimwipe is a registered trademark of Kimberly Clark Corporation Irving TX USA Pentium is a registered trademark of Intel Corporation Santa Clara CA USA Windows and Excel are registered trademarks of Microsoft Corporation Redmond WA USA Copyright Information The contents of this document are the property of Tecan Austria GmbH and are not to be copied ...

Страница 31: ...hlight important information or to warn the user of a potentially dangerous situation Note Gives helpful information Caution Indicates a possibility of product damage or data loss if instructions are not followed Warning Carefully read and follow the instructions provided in this document before using the software or using the product Symbols Manufactured by ...

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