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NanoPhotometer

® 

P-Class

 User Manual 

 

 
Version 2.1 

 

Page 21 / 70 

Results Screen 
 
 

 

 

  Results Screen 
 

Step 10

 

Apply/insert  the  reference  sample.  Press 

Blank

  Key.  This 

will be used for all subsequent samples until changed. 

Step 11

 

Apply/insert sample and press 

Sample

 

. This measures 

at  both  260  and  280  nm  wavelengths  and  displays  the 
result.  Protein  concentration  is  calculated  (corrected  by 
background wavelength value if selected). 

Step 12

 

If  the  absorbance  value  of  the  sample  is  not  in  the  linear 
range  a  “Warning  message”  will  pop  up  and  “Instruction” 
will be displayed in the top left corner of the result screen. 
Please  refer  to  3.2  Software  instructions/important 
information on page 11 for further information. 

Step 13

 

Repeat for all samples.  

Step 14

 

Press 

Menu/Options

 to display available Options which are 

described on page 8. 

Step 15

 

Press 

Escape

 

 and confirm with 

Yes

 

 to return to the 

Protein

 folder 

 
To change parameters, print or save methods press the Menu/Options button. The options menu will be opened. For 
further explanation please see 2.3 Keypad and display on page 6 (P 300) and 7 (P330 / P 360). 
 

Summary of Contents for NanoPhotometer

Page 1: ...telephone 49 89 726 3718 0 Fax 49 89 726 3718 54 Email info implen de www implen de NanoPhotometer P Class User Manual P 300 P 330 P 360 Version 2 1 P 360 P 300 P 330...

Page 2: ...e 1995 5 EC Radio and Telecommunications Terminal Equipment Directive instruments fitted with Bluetooth accessory only Standards to which conformity is declared where relevant are as follows EN 61010...

Page 3: ...in Determination 19 4 2 1 General Information 19 4 2 2 Protein UV Method 20 4 2 3 Protein UV Dye Method 22 7 3 4 BCA Assay 24 7 3 5 Bradford Assay 27 7 3 6 Lowry Assay 30 7 3 7 Biuret Assay 33 4 3 Bac...

Page 4: ...shooting 64 9 ACCESSORIES 64 10 SPECIFICATION AND WARRANTY 65 11 SOLVENT COMPATIBILITY 66 12 APPENDIX 67 12 1 Nucleic acid quantification 67 12 2 Nucleic acid fluorescent dye incorporation 67 12 3 Pro...

Page 5: ...has been established 2 3 hours Maximum relative humidity of 80 up to 31 C decreasing linearly to 50 at 40 C The instrument must be placed on a stable level bench or table that can take its weight 4 5...

Page 6: ...or in native NanoPhotometer P Class Software format for later access see also NanoPhotometer P Class PVC Installation and User Manual Alternatively results may be saved on a SD Memory Card P300 only o...

Page 7: ...levant section of the NanoPhotometer P Class User Manual Alphanumeric keys Use these to enter parameters and to write text descriptions where appropriate or required Use repeated key presses to cycle...

Page 8: ...here appropriate or required Use repeated key presses to cycle through lower case number and upper case Leave for 1 second before entering next character Use C button to backspace and 1 to enter a spa...

Page 9: ...art from 8 Save the parameters as a method 9 Open printer settings possibility to change the printer settings within the method as described in 7 3 Output Options Printer Exit Options by pressing Esca...

Page 10: ...Photometer P Class Submicroliter Cell into the cell holder with the cell windows facing the light beam We recommend facing the Implen logo to the front The light beam is directed from RIGHT to LEFT as...

Page 11: ...pplications and Cuvette Applications The procedure is as follows Exemplary Parameter Screen Parameter Screen Step 1 Press 1 to select NanoVolume Applications folder Step 2 Press 1 to select Nucleic Ac...

Page 12: ...id 100 Lid 100 Concentration too low Do you want to change the lid factor Please change to lid 50 and press sample automatic change of lid factor lid 100 to lid 50 in the software for calculation No c...

Page 13: ...e 10 mm pathlength Dilution factor and cell pathlength can be entered Nucleic Acid Purity Checks Nucleic acids extracted from cells are accompanied by protein and extensive purification is required to...

Page 14: ...cent dye incorporation To determine the dye incorporation rate the absorbance reading at the wavelength reported for maximum absorbance of the fluorescence dye is used The corresponding extinction coe...

Page 15: ...ng the left and right arrows Options g ml ng l g l Step 8 Enter the Factor using the keypad numbers Default value is 50 for dsDNA 37 for ssDNA and 40 for RNA range is 0 01 to 9 999 Step 9 Press OK to...

Page 16: ...ction table denoting the ratios of the 4 bases according to the oligo sequence Enter the proportions of bases present using the keypad numbers and up and down arrows to move between boxes Default is 1...

Page 17: ...to select one of the dye incorporation methods Step 4 Using the NanoVolume Applications select the Lid Factor as described in the Average Detection Range Sheet and under 3 2 Step 5 Select Dilution Fac...

Page 18: ...f the absorbance value of the sample is not in the linear range a Warning message will pop up and Instruction will be displayed in the top left corner of the result screen Please refer to 3 2 Software...

Page 19: ...cations and Cuvette Applications To determine the degree of labelling the absorbance reading at the wavelength reported for maximum absorbance of the fluorescence dye is used The corresponding extinct...

Page 20: ...e keypad numbers Range 0 01 to 9 999 Press OK to calculate the dilution factor and return to the Parameters screen OR press Cancel to cancel the selections and return to the Parameters screen Step 5 S...

Page 21: ...alue of the sample is not in the linear range a Warning message will pop up and Instruction will be displayed in the top left corner of the result screen Please refer to 3 2 Software instructions impo...

Page 22: ...ein BSA default Serum Albumin mouse Serum Albumin human IgG Lysozyme Custom or OD 1 Step 8 If using Custom Protein there are two possibilities to enter the correct factors see also page 18 protein UV...

Page 23: ...ulated Step 16 If the absorbance value of the sample is not in the linear range a Warning message will pop up and Instruction will be displayed in the top left corner of the result screen Please refer...

Page 24: ...significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen parameters OR Cancel Step 7 Pres...

Page 25: ...measured press OK to accept the calibration and go to the Results screen see below OR press Back to cancel selections and return to the Standards screen Calibration Screen replicates on Step 17 This s...

Page 26: ...id unintended escaping the application NanoPhotometer P 300 Options select using key pad numbers NanoPhotometer P 330 P360 Menu select using key pad numbers Options select using key pad numbers 1 Retu...

Page 27: ...to 5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen parameters OR Cancel Step...

Page 28: ...ured press OK to accept the calibration and go to the Results screen see below OR press Back to cancel selections and return to the Standards screen Calibration Screen replicates on Step 17 This shows...

Page 29: ...oid unintended escaping the application NanoPhotometer P 300 Options select using key pad numbers NanoPhotometer P 330 P360 Menu select using key pad numbers Options select using key pad numbers 1 Ret...

Page 30: ...5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen parameters OR Cancel Step 7 P...

Page 31: ...ured press OK to accept the calibration and go to the Results screen see below OR press Back to cancel selections and return to the Standards screen Calibration Screen replicates on Step 17 This shows...

Page 32: ...unintended escaping the application NanoPhotometer P 300 Options select using key pad numbers NanoPhotometer P 330 P360 Menu select using key pad numbers Options select using key pad numbers 1 Return...

Page 33: ...5 significant figures regardless of how many decimal points are selected so 98768 2 will display as 98768 even with 1 decimal point selected Press OK to store the chosen parameters OR Cancel Step 7 P...

Page 34: ...ured press OK to accept the calibration and go to the Results screen see below OR press Back to cancel selections and return to the Standards screen Calibration Screen replicates on Step 17 This shows...

Page 35: ...oid unintended escaping the application NanoPhotometer P 300 Options select using key pad numbers NanoPhotometer P 330 P360 Menu select using key pad numbers Options select using key pad numbers 1 Ret...

Page 36: ...t geometry of this slit and the monochromator optics Different spectrophotometer types therefore give different responses for the same turbid sample to compare results they must be normalized using ca...

Page 37: ...ght arrows Options are 1 000 or 1 000 000 Factor and Multiplier define the conversion of the measured OD to the number of cells per millilitre e g 1 OD 600 5 x 108 cells ml Step 8 Press OK to enter th...

Page 38: ...ed on a user programmed curve 6 Absorbance or T transmission at up to 5 user defined wavelengths 7 Ratio of absorbance values at two user specified wavelengths Menu Options Within each function the us...

Page 39: ...ep 5 To enter the results screen with the selected parameters press OK OR cancel the selections and return to the Functions folder by pressing Cancel Results Screen Step 6 Insert the reference sample...

Page 40: ...d enter name 9 Open printer settings possibility to change the printer settings within the method as described in 7 3 Output Options Printer Exit Options by pressing Escape or wait Menu select using k...

Page 41: ...utton to delete the last digit entered Step 6 if Standard is selected Enter the concentration using keypad numbers Range 0 01 to 9 999 Use the C button to delete the last digit entered Step 7 Units Th...

Page 42: ...pad numbers 1 Return to parameters screen 2 Print result via selected method 3 Toggles on off displaying a graph of wavescan 20 nm from selected wavelength 4 Return to Run Standard screen 7 Sample nu...

Page 43: ...ncel Measurement Screen Step 7 Insert the reference sample Press Blank key This will be used for all subsequent samples until changed Step 8 Insert sample and press Sample Step 9 Repeat for all sample...

Page 44: ...settings within the method as described in 7 3 Output Options Printer Exit Menu Options by pressing Escape or wait Peak Detection Shortcut button 4 Auto Detect Peaks Turns on and off the automatic pe...

Page 45: ...aph Scale Shortcut button 6 This enables the user to set up a defined graph by defining the limits in either or both of the x and y axes Zoom mode This sets up the operation of the Zoom keys up and do...

Page 46: ...minutes Step 5 Duration Enter the time in minutes over which measurements are taken This can be a maximum of 60 minutes Step 6 Interval Enter the interval time in seconds between measurements using th...

Page 47: ...Query needs confirmation to avoid unintended escaping the application Menu select using key pad numbers 1 Return to parameter screen 2 Transfer the results via selected Output Options Printer setting...

Page 48: ...e Menu Options key and then use the left right arrows g ml g l pmol l mg ml mmol l mol l g l mg l g l U l ppm ppb conc or none These units can also be edited once OK is pressed This screen also allows...

Page 49: ...results and the fitted curve as the measurements are input Step 16 Use the up and down arrows to select a standard to be repeated if a poor reading has been obtained Use C to clear the previous readi...

Page 50: ...ll subsequent samples until changed Step 28 Insert the sample and press Sample The concentration of the sample is taken and displayed Step 29 Repeat for all samples Step 30 Press Menu Options to displ...

Page 51: ...bove and repeat for the number of wavelengths selected up to 5 Step 7 Press OK to enter the results screen OR press Cancel to return to the Functions folder Results Screen Step 8 Insert the reference...

Page 52: ...tore in User Methods 1 9 press the down arrow and enter name 9 Open printer settings possibility to change the printer settings within the method as described in 7 3 Output Options Printer Exit Option...

Page 53: ...ction is On Enter the third Wavelength from which the background correction will be obtained Step 7 Press Next to enter the next screen OR press Cancel to return to the Functions folder Step 8 Dilutio...

Page 54: ...ions which are described below Step 18 Press Escape and confirm with Yes to return to the Functions folder Query needs confirmation to avoid unintended escaping the application Menu select using key p...

Page 55: ...ws 4 Press OK to lock the method OR Cancel to return to the User Methods folder Unlock Method 1 Press 3 to select Unlock Folder 2 Select the method to be unlocked using the left and right arrows 3 Ent...

Page 56: ...thod 3 Press OK to delete the method OR Cancel to return to User Methods folder 7 UTILITIES Survey of the available Utilities Key pad number Description 1 Set correct time and date 2 Select preferred...

Page 57: ...s folder OR press Cancel to return to the Utilities folder without storing the settings 7 3 Output Options Printer 7 3 1 NanoPhotometer P 300 The NanoPhotometer P 300 offers a selection of data output...

Page 58: ...ease proceed to Folder 5 Utilities press 3 Output Options to set the favoured reporting settings Data output to Built in printer Printing on the built in printer is possible independent from the Outpu...

Page 59: ...urn to the Utilities folder without storing the settings If Computer USB is selected all measured data are sent automatically to the PC The file is closed and saved once the method is left by pressing...

Page 60: ...urs at night or off Step 5 Select Baseline Compensation to improve value stability and to overcome background effects Step 6 Press OK to store the settings and return to the Utilities folder OR press...

Page 61: ...ing GLP GMP calibration certification using filters traceable to international standards during production and quality control certified engineers and calibrated test equipment approved to ISO 9001 st...

Page 62: ...isinfection Desinfection or cleaning only by wiping no spraying Use no solvents or aggressive chemicals Approved disinfectant solutions Apesin disinfection spray Tana Chemie GmbH Incidin Liquid and In...

Page 63: ...n No light on Reference channel Submicroliter cell or cuvette in the cell holder by switching on the instrument Cell holder has been removed from the instrument and placed back in the wrong position R...

Page 64: ...of paper normal sound no error message Thermal Printer paper was probably put the wrong way check correct position of the printer paper Thermal printer automatic roller stops Please contact the Implen...

Page 65: ...n factors 5 10 50 100 and 250 Other technical data Vortexer 2 800 rpm tube size up to 2 0 ml Cuvette storage capacity for eight 10 mm cells Photometric mode Abs T concentration scan ratio multi wavele...

Page 66: ...nzene butanol carbon tetrachloride chloroform ethanol ether HEPES hexane isopropanol MES methanol methylenchlorid MOPS phenol up to 1 n propanol toluene buffers containing phosphate or chloride salts...

Page 67: ...on on and off Calculation of the fluorescence nucleic acid concentration c nuc Abs 260 Abs 320 factor nuc lid factor dilution factor With dye correction c nuc Abs 260 Abs 320 CF dye Abs max dye Abs 32...

Page 68: ...cm 1 The following dye types and parameters are pre programmed in the NanoPhotometer P Class Dye Type Absorption maximum Dye nm Dye dependent extinction coefficient dye Dye dependent correction factor...

Page 69: ...incorporation To determine the protein concentration and the dye concentration after labelling a modification of the Lambert Beer equation is used Background correction is always calculated possibili...

Page 70: ...ependent molar extinction coefficient M 1 cm 1 The following dye types and parameters are pre programmed in the NanoPhotometer P Class Dye Type Absorption maximum Dyes nm Dye dependent extinction coef...

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