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Appendix C - Calculations

Thermo Electron Corporation

C-4

Calculation

Entry Parameters

Formula

Displayed Units

Calculation of Tm: • # units A, # units T,

T

m

= 78

°

C + 16.6log ((Na

+

)/ 

°

C

RNA-RNA hybrids

# units G, # units C

(1+0.7(Na

+

)) + 0.7(%GC) – 

• M = molarity of cation

500/L – P – 0.35(%formamide)

• Fraction GC = fraction 

of G and C

• %form = %formamide 

in the sample

• L = # of base pairs
• P = % mismatching

Conversion from

• 

µ

g/mL and molecular

pmol/

µ

L

=   

µ

g/mL x 1000

pmol/

µ

L

weight from Oligo (calc                       DNA Mol. Wt.
factor) test

Summary of Contents for 335904

Page 1: ...BioMate 3 Series Spectrophotometers Operator s Manual ...

Page 2: ......

Page 3: ...Figure 1 BioMate 3 spectrophotometer Figure 2 Back panel of the spectrophotometer Key c A C power connector d RS232C port ...

Page 4: ...paper roll holders for the internal printer 335988 Figure 4 Keypad of the BioMate 3 spectrophotometer Key c Cell position keys d Function keys Key c Paper roll holder d Icon for paper direction e Paper entry slot f Finger tab ...

Page 5: ...Figure 5 Installing the Single Position Cell Holder 335916 Figure 6 Installing the printer 335988 Key c Captive screw on lamp door d Removing door from hinge e Hinge f Connecting wires to printer ...

Page 6: ...Figure 7 Removing the cover of the fuse compartment Figure 8 Removing the fuse holder Figure 9 Removing and replacing the fuses ...

Page 7: ...netic Compatibility Test Standards IEC 61326 1 1998 Electrical Equipment for Measurement Control and Laboratory use EMC Requirements Class A Limits IEC 61000 4 2 1999 Electrostatic Discharge Immunity Test Test level 4KV Air Discharge and 4 KV Contact Discharge IEC 61000 4 3 1998 Radiated Radio Frequency Electromagnetic Field Immunity Test Test level 3V m IEC 61000 4 4 1995 Electrical Fast Transien...

Page 8: ......

Page 9: ...es of merchantability and fitness for a particular purpose Copyright 2003 by Thermo Electron Scientific Instruments Corporation Madison WI 53711 Printed in the United States of America All world rights reserved No part of this publication may be stored in a retrieval system transmitted or reproduced in any way including but not limited to photocopy photograph magnetic or other record without the p...

Page 10: ...rvice personnel This warranty does not apply to equipment and accessories that have been modified or tampered with in any way misused or damaged by accident neglect or conditions beyond Thermo Electron Scientific Instruments Corporation s control This warranty does not apply to lamps glassware and similar expendable components However such parts and components may be warranted by their manufacture...

Page 11: ...orporation spectrophotometer Through the security section you can lock test setups test parameters so that they may not be altered The password also allows you to remove the security so that you may edit the test parameters Please refer to the appropriate section in this Operator s Manual for more information on locking a test PASSWORD 4 3 6 3 7 9 7 ...

Page 12: ...Thermo Electron Corporation iv ...

Page 13: ...c acid measurements 2 2 DNA 260 280 and DNA 260 230 2 3 DNA with Scan 260 280 and DNA with Scan 260 230 2 4 dsDNA ssDNA RNA and Oligos entered factor Direct or UV Measurements 2 5 Protein Measurements 2 8 Bradford standard micro Lowry standard micro BCA standard micro and Biuret measurements 2 8 Direct UV 280 and Direct UV 205 2 9 Warburg Christian 2 10 Cell Growth 2 11 Setting test parameters 2 1...

Page 14: ...rs for a standard curve 3 12 Measuring the standards for a standard curve 3 12 Measuring unknowns 3 13 Editing a standard curve 3 13 Absorbance Ratio 3 13 Recalling a test 3 14 Setting up test parameters 3 15 Measuring unknowns 3 15 Absorbance Difference 3 16 Recalling a test 3 16 Setting up test parameters 3 16 Measuring unknowns 3 16 Kinetics 3 17 Recalling a test 3 17 Setting up test parameters...

Page 15: ...3 Deleting standards 4 4 Running the test 4 4 Noise Measurement 4 4 Stray Light 4 5 Running the test 4 5 Internal Printer Test 4 5 RS232C Test 4 6 CHAPTER 5 Connecting Using Accessories General information 5 1 Cell holders cell holder accessories 5 1 Changing cell holders 5 2 Internal printer 5 3 Installing the internal printer 5 3 Loading paper in the internal printer 5 3 External computers 5 3 C...

Page 16: ...Table of Contents Appendix A Specifications Appendix B Parameters Appendix C Calculations Index Thermo Electron Corporation viii ...

Page 17: ...ent by pressing the power switch to ON 1 ON 0 OFF The power on sequence will appear on the display and the instrument will go through its self diagnostics The instrument performs these diagnostics in the sequence shown a Logo b Initializing c Calibrating filter wheel d Finding zero order e Finding energy peak f Calibrating grating Setting utility parameters You should set up some of the utility pa...

Page 18: ...en displayed press the arrow keys to highlight Standby and press ENTER Press the arrow keys to highlight the length of time you want the instrument will wait before entering standby mode and press ENTER Setting baseline expiration time If you will be performing scans on your samples you can set a time limit for a collected baseline To set the baseline expiration time With the Utility screen displa...

Page 19: ... Quartz From 190 to 230nm up to 1100nm Disposable Refer to manufacturer s specifications and ensure that you work within the recommended range Test tubes vs cuvettes Square cuvettes that are carefully matched see Correcting for cell variability below yield very precise results Matched test tubes when properly handled can show as little as 1 2 deviation between readings The pathlength of test tubes...

Page 20: ...the light beam in the spectrophotometer The specifications for the sample compartment including the dimensions of the beam are Z dimension Square cuvettes Test tubes 8 5mm Beam size 2mm wide x 7mm high Setting Up the Instrument Thermo Electron Corporation 1 4 ...

Page 21: ...ndows list multiple options for a parameter or display messages You can press the arrow keys to select the value you want then press ENTER to select it and move to another parameter Cursor selections are used on graphical displays and the character list you use to name tests and files On graphical displays the cursor appears as a vertical line on the character list the cursor highlights the charac...

Page 22: ...e test you selected will be displayed Note At this point you can power down the instrument and then power it back up When it starts up again the parameter screen for the selected test will be displayed Setting up a multiple test SmartStart 1 Press the Stored Tests key on the spectrophotometer keypad 2 Scroll down through the list until the first appropriate test is highlighted 3 Press Select Tests...

Page 23: ...BioMate Tests screen displayed move the arrow keys to highlight Nucleic Acid Tests and press ENTER A list of nucleic acid test appears Move the arrow keys to highlight DNA 260 280 The DNA 260 280 parameter screen appears Note The following screens show the parameters for the DNA 260 280 test For the DNA 260 230 test Wavelength 2 is set to 230nm Note If Cell Correction is ON you must run the Setup ...

Page 24: ...and 230nm Refer to Appendix B for a description of the parameters and Appendix C for the default values To get started with the BioMate Tests screen displayed move the arrow keys to highlight Nucleic Acid Tests and press ENTER A list of nucleic acid test appears Move the arrow keys to highlight DNA with Scan 260 280 The DNA with Scan 260 280 parameter screen appears Note The following screens show...

Page 25: ...between tabular and graphical displays press Graph or Tabular Note You may need to use the up and down arrow keys to view all the data for the screen dsDNA ssDNA RNA and Oligos entered factor Direct or UV Measurements These test measurements are all set up and run using the same types of test parameters Refer to Appendix B for a description of the parameters and Appendix C for the default values T...

Page 26: ...d with a 6 Position Cell Holder you can place up to five samples in the cell holder 3 Press Measure Blank to measure the blank If your instrument is equipped with a 6 Position Cell Holder it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position 4 Press Measure Sample to start the measurement If your instrument is equip...

Page 27: ...ress ENTER to start the measurements The instrument automatically measures the blank first then measures the unknowns and displays absorbance oligo concentration in µg mL and pmol µL Measuring unknowns manually using Manual 6 or Single Cell Positioner 1 With the Oligos calc factor screen displayed press Run Test 2 Place the blank and unknown in the correct cell positions If your instrument is equi...

Page 28: ...for the Bradford Standard test All the other protein standard curve methods work in a similar fashion Note If Cell Correction is ON you must run the Setup Correction program before you can access the Run Test or Measure Samples keys Setting up test parameters for a standard curve 1 With the Bradford Standard setup screen displayed use the arrow keys to highlight the name of the parameter you want ...

Page 29: ...to start the measurements The instrument automatically measures the blank first then measures the unknowns and displays the absorbance and concentration of each unknown Measuring unknowns manually using Manual 6 or Single Cell Positioner 1 With the Standards screen displayed press Run Test 2 Place the blank and unknown in the correct cell positions If your instrument is equipped with a 6 Position ...

Page 30: ... Position Cell Holder it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position 4 Press Measure Sample to start the measurement If your instrument is equipped with a 6 Position Cell Holder press the cell position buttons on the keypad to re position the cell holder and measure the rest of the unknowns manually Warburg C...

Page 31: ...progress of cell growth in a sample The instrument does not perform any calculations or graphing for the data To get started move the arrow keys on the BioMate Tests screen to highlight Cell Growth and press ENTER The Cell Growth setup screen appears Note If Cell Correction is ON you must run the Setup Correction program before you can access the Run Test or Measure Samples keys Setting up test pa...

Page 32: ...RNA and RNA RNA hybrids Refer to Appendix B for a description of the parameters and Appendix C for the default values To get started move the arrow keys on the BioMate Tests screen to highlight Oligo Calculator and press ENTER The Oligos screen appears Using the oligo calculator 1 With the Oligos screen displayed press Base Sequence The Base Sequence identification screen appears 2 Use the arrow k...

Page 33: ... select the characters for the filename from a character list 1 After setting up the values for the test parameters press Save Test The Create Test Name screen appears You can use this screen to Delete the name of a test Delete a character in the name of a test Add a character to the name of a test Accept the name of a test 2 Use the arrow keys to highlight the first character you want to use for ...

Page 34: ...d all the characters for the name 4 Press Accept Name to accept the name and return to the previous screen The name of the new custom unit appears on the list of basic units Using the SmartStart feature The SmartStart feature enables you to select the test methods you use most frequently and have them appear when you start up your instrument If your laboratory runs only a single test you can use t...

Page 35: ...not active in the Main Basic Absorbance T Basic Concentration screen Note The Cell Correction feature is active only when the 6 Position Sample Holder is set to either Auto 6 or Auto 3 The feature is not active when the cell holder is set to 1 Cell Platform or Manual 6 nor when the Single Cell Holder is installed Every test setup screen provides access to the cell correction program Before running...

Page 36: ...te nms mode first specify the wavelengths using the procedures which follow and then the Cell Correction program The Cell Correction program will measure the other cells against the blank and will record store and date the measurements From these measurements the Cell Correction program establishes the required correction factors which then are automatically applied during all subsequent tests if ...

Page 37: ... not function when you select Single Cell Platform even if you have a 6 Position Cell Holder installed in the sample compartment Manual 6 You can take up to six measurements without changing the samples in the cell holder using the cell position buttons on the keypad to advance the cell holder to the appropriate position for the next measurement You place the blank in the blank position and your s...

Page 38: ...ell holder If your instrument has a 6 Position Cell Holder be sure to place the blank in the B position 2 If you want to enter an absorbance or transmittance value for the blank press a number key and enter the concentration in the Entry field 3 Press Measure Blank to measure the blank When the instrument is finished measuring the absorbance of the blank the message disappears Measuring unknowns 1...

Page 39: ...on for the blank press a number key and enter the concentration in the Entry field 3 Press Measure Blank to measure the blank When the instrument is finished measuring the absorbance of the blank the message disappears Measuring a standard 1 If necessary press Change Mode to switch to the Concentration with Standard mode 2 If your instrument is equipped with the 6 Position Cell Holder place the st...

Page 40: ...e measurement appears on the display If your instrument is equipped with the Single Cell Holder Platform remove the blank and place the unknown in the cell holder The measurement appears on the display Advanced A T C Absorbance Transmittance measurements When you use the Advanced A T C program for absorbance or transmittance measurements you can perform these tasks Select the measurement mode you ...

Page 41: ...easurement screen appears prompting you to place your samples in the cell holder 2 Place the blank and unknowns in the cell holder If your instrument is equipped with a 6 Position Cell Holder be sure to place the blank in the B position You can place up to five samples in the cell holder 3 Press Measure Blank to measure the blank If your instrument is equipped with a 6 Position Cell Holder it auto...

Page 42: ...tandard automatically using Auto 6 or Auto 3 1 With the Advanced A T C screen displayed and the Measurement Mode set to Conc Std press Run Standard The Measure Standard screen appears 2 Use the number keys to enter the concentration value of the standard and press ENTER 3 Press Measure Standard 4 Place the blank and standard in the correct cell positions 5 Press ENTER to measure the blank and the ...

Page 43: ...using Manual 6 or Single Cell Platform 1 With the Advanced A T C screen displayed and the Measurement Mode set to Conc Std or Conc Factor press Run Test The Advanced A T C measurement screen appears 2 Place the blank and unknown in the correct cell position If your instrument is equipped with a 6 Position Cell Holder you can place up to five samples in the cell holder 3 Press Measure Blank to meas...

Page 44: ... all the concentration values the Standards screen appears 4 Press Measure Standards to measure the blank and standards The instrument automatically measures the blank first then measures the standards When the instrument has measured all the standards the Standards screen appears showing the absorbance of each standard along with the slope intercept and correlation coefficient of the standard cur...

Page 45: ...pears showing the absorbance and concentration of each unknown Measuring unknowns manually using Manual 6 or Single Cell Platform 1 With the Standards screen displayed press Run Test The Samples screen appears 2 Place the blank and unknowns in the correct cell positions 3 Press Measure Blank to measure the blank If your instrument is equipped with a 6 Position Cell Holder it automatically moves to...

Page 46: ...yed on your screen press Edit Standards 2 Use the arrow keys to highlight Clear Measurements and press ENTER All the absorbance measurements will be removed from the screen To reset standards 1 With the standard curve displayed on your screen press Reset Standards and press ENTER 2 Use the arrow keys to highlight Reset Standards and press ENTER All the standards and measurements will be removed To...

Page 47: ... Test to measure a blank or unknowns Measuring unknowns Measuring unknowns automatically using Auto 6 or Auto 3 1 Place the blank and unknowns in the correct cell positions 2 Press ENTER to start the measurement The instrument automatically measures the blank first then measures the unknowns and displays the sample measurements on the screen Measuring unknowns manually using Manual 6 or Single Cel...

Page 48: ...ram Save a test View the list of stored tests Measure a blank Measure unknowns Setting up test parameters 1 With the Absorbance Difference screen displayed use the arrow keys to highlight the name of the parameter you want to set 2 When the parameters are set you can press Save Test to save the test or Run Test to measure a blank or unknowns Measuring unknowns Measuring unknowns automatically usin...

Page 49: ...run When you set up your test parameters be sure to select the interval time and total run time accordingly To get started press the TEST key on the keypad When the Test Types screen appears highlight Kinetics and press ENTER Recalling a test 1 With the Kinetics screen displayed press Stored Tests A list of stored tests appears 2 Use the arrow keys to highlight the name of the test you want to rec...

Page 50: ...e When you are working with graphical kinetics results you need to press Edit Graph before you can rescale and recalculate You can modify the scale of your kinetics data plot in two ways automatically or manually When you select Auto Scale the instrument automatically scales the X and Y axes so all the data appears on the plot When you select Manual Scale you select specific minimum and maximum va...

Page 51: ... with tabular kinetics results you need to press Edit Data before you can rescale and recalculate After collecting your kinetics data you can use all the data for the rate calculation or you can select specific start and end times Whenever you modify the start and end times or select all the data the instrument automatically recalculates and displays the new reaction rate and result When the edit ...

Page 52: ...ests appears 2 Use the arrow keys to highlight the name of the test you want to recall and press ENTER The parameters for the selected test appear on the screen From this screen you can Set up test parameters Set up Cell Correction program Save a test View the list of stored tests Collect a baseline Scan unknowns Setting up test parameters 1 With the Survey Scan Scanning screen displayed use the a...

Page 53: ... to press Edit Graph before you can perform other functions on the scan data When the edit graph screen appears you can Rescale the graph Perform mathematical operations on the graph Rescaling graphical scan data You can modify the scale of your scan data plot in two ways automatically or manually When you select Auto Scale the instrument automatically scales the X and Y axes so all the data appea...

Page 54: ...the appropriate function key Min Y Max Y Min X Max X to accept it The instrument redraws the plot using the minimum and maximum values you have entered To use the Auto Scale function 1 With your scan data displayed on the edit scale screen press Auto Scale The instrument automatically adjusts the minimum and maximum values for the X and Y axes so all the data appears on the plot Performing calcula...

Page 55: ...the desired wavelength value The instrument calculates the area under the curve for the selected wavelengths 4 Continue selecting the other wavelengths by pressing Next Cursor to activate the next cursor line Select the wavelength by positioning the cursor with the Cursor and Cursor keys 5 Press Set Options to access the set options window 6 Use the up and down arrows to highlight Factor Enter the...

Page 56: ...rted press the TEST key on the keypad When the Test Types screen appears highlight 3 Point Net and press ENTER Recalling a test 1 With the 3 Point Net screen displayed press Stored Tests A list of stored tests appears 2 Use the arrow keys to highlight the name of the test you want to recall and press ENTER The parameters for the selected test appear on the screen From this screen you can Set up te...

Page 57: ...ll position 4 Press Measure Sample to measure the unknowns The sample measurement appears on the screen If your instrument is equipped with a 6 Position Cell Holder press the cell position buttons on the keypad to reposition the cell holder and measure the rest of the unknowns manually Multiple Wavelengths When you use the Multiple Wavelengths program you can perform these tasks Recall a test OR s...

Page 58: ... nm 4 Enter the values for the wavelength and factor and press ENTER 5 When the values are correct press Add nm 6 Continue until you have entered all the wavelengths and factors Deleting wavelengths factors 1 With the Multiple Wavelengths screen displayed use the arrow keys to highlight Set nms The wavelength screen appears listing the wavelengths and factors specified for the measurements Note If...

Page 59: ...older it automatically moves to the B position to measure the blank When it completes the measurement it returns to its previous cell position 4 With the list of wavelengths and factors displayed press Measure Samples to measure the unknowns The instrument measures the absorbance at each wavelength and displays it on the screen If you have set the measurement mode to Concentration Factor the instr...

Page 60: ...Using General Tests Software Thermo Electron Corporation 3 28 ...

Page 61: ... fail If a test continues to fail after you have tried all the recommendations in the list follow the troubleshooting list for the test being run included with the description of each test Make sure you are following the instructions for the test properly Make sure filters and standards are clean Make sure the sample compartment door is closed while the test is running Make sure the sample compart...

Page 62: ...rtificate of calibration included with your standards Note Only the middle wavelength of the SPECTRONIC Standards wavelength standard is certified The wavelength accuracy test passes or fails based on this wavelength alone The other two wavelengths may be used for long term repeatability testing The initial target values may be determined by running the wavelength accuracy test once and taking not...

Page 63: ... and compares the results with specified tolerances The absorbances and tolerances are preset but you should change them to the values on the report of calibration included with your standards Note You can display the tolerances for this test in either absorbance or transmittance 1 With the Performance Validation screen displayed use the arrow keys to highlight Photometric Accuracy 2 Press ENTER T...

Page 64: ... the test is failing consistently Make sure you follow the guidelines provided with the standard reference materials Contact technical support for more troubleshooting advice Noise Measurement This test measures the amount of noise peak to peak at specified wavelengths UV Vis instruments measures noise at 230 and 340nm All parameters are preset and cannot be changed When running the noise test rem...

Page 65: ...u may use other filters but they must have 0 1 T at the wavelength of interest 1 With the Performance Validation screen displayed use the arrow keys to highlight Stray Light 2 Press ENTER The Stray Light screen appears Running the test 1 With the Stray Light screen displayed make sure that the wavelengths and tolerances are set correctly 2 Press Start Test to run the test The results of the test a...

Page 66: ...test allows you to verify that the RS232C port is functional To run the test you will need to have the RS232 Test Plug 336035 attached to the RS232C port on the back panel of the instrument Running the RS232C test takes approximately five minutes 1 Verify that the RS232 Test Plug is securely attached to the RS232C port on the back panel of the instrument 2 With the Performance Validation screen di...

Page 67: ...r holds three or 335916 Single Cell Holder Platform holds one 335079 Thermal Block Controls the temperature of one 10mm pathlength square cuvette within the range of 20 to 100ºC fits on 335927 6 Position Cell Holder holds three or 335916 Single Cell Holder Platform holds one requires external water bath and 335921 Accessory Door for routing circulating water 336028 Single Cell Holder Accommodates ...

Page 68: ...u will not be able to tighten the screws 3 Take a blank Measurement to ensure that the instrument detects that you have installed the Single Cell Platform Installing accessory cell holders 1 Make sure that you have the correct cell holder installed If you want to use 100mm longpath cells you must install the Single Position Cell Holder 2 Install the desired accessory cell holders by tightening the...

Page 69: ...he plastic door 9 Use the clip on the hinge to secure the wires 10 Install the printer door by sliding it back onto the hinge 3 Figure 6 11 Close the lamp door 12 Tighten the captive screw 1 Figure 6 on the printer door to hold it securely in place 13 Load the paper into the printer see instructions in Setting up the instrument Loading the paper in the internal printer For instructions on loading ...

Page 70: ...Thermo Electron Corporation 5 4 Connecting Using Accessories ...

Page 71: ...ge to the instrument If concentrated acids or bases or any hydrocarbon materials are spilled on the instrument be sure to clean up the affected area immediately Use water alcohol or other common laboratory cleaning agents to clean the keyboard It is recommended that you clean spills off the keyboard as soon as they occur Cleaning Cleaning and maintenance of cells Cleaning of cells both inside and ...

Page 72: ...Be sure to remove all fingerprints Changing the fuse The fuse is located in the power entry module located at the center of the back panel of the instrument Figure 2 120VAC 2 5A Slo Blo 240VAC 1 25A Slo Blo 2 required See Replacement parts on page 5 3 for a list of replacement part numbers WARNING The instrument fuse must be replaced with the same type and rating fuse WARNING If the fuse fails rep...

Page 73: ...36028 single cell holders installed 335989 Internal Printer Assembly 333152 Sensitized Paper 100 sheets for SPECTRONIC Standards 335965 Replacement Dust Cover 335954 Printer Paper Package of 5 rolls 335904 10001 BioMate 3 Operator s Manual 335901 10020 Service Manual includes printed service manual and CD ROM 335901 10040 BioMate 3 Service Manual includes CD ROM only 335971 Replacement silicone tu...

Page 74: ...Performing Maintenance Procedures Thermo Electron Corporation 6 4 ...

Page 75: ...hour Stray Light4 0 1 T at 220 and 340nm Standard Interface Bi directional RS232C Standard Cell Holder 1 position and automatic 6 position cell holder Keyboard Membrane keypad Software DNA ratio concentration with or without scanning Direct protein at 280nm Protein standard curves Cell growth Oligo calculator absorptivity molecular weight factor and theoretical Tm Absorbance Transmittance Concentr...

Page 76: ...nds moisture per pound of dry air Allow instrument to adjust to room temperature for 24 hours after taking it out of storage Temperature should be maintained at 4 F Relative humidity should be maintained to 5 Altitude From below sea level to 2000 meters 6562 feet For indoor use only Installation Category II Pollution Degree 2 BioMate 3 Dimensions 330W x 410D x 235H mm 13 x 16 x 9 Weight 8 6 Kg 19l...

Page 77: ...uvetts used to hold and measure samples Cell Pos Position placed in light path set by selection of sample positioner Concentration Sets concentration value Correction Mode Mode for cell correction collection Cursor position on Indicates which option on the Test Types screen should be highlighted Test Types menus Cursor position on Indicates which option on the Utility screen should be highlighted ...

Page 78: ...protect stored tests from accidental deletion or alteration Low High Limits Lowest highest acceptable results outside of which the result is flagged as Low or High Measurement Mode The type of photometric data reported for a measurement Molarity of cation Molarity of Na in incubation mixture Number of Bases Number of bases in the oligonucleotide Number of Matched Number of cuvettes that will be ru...

Page 79: ...to generate standard curve for the test Standby Time since last keystroke or instrument activity powers down unit to minimum energy required to keep lamp in warm up status Statistics Turns statistics function on or off if ON calculates average and Std Dev of results Statistics registers are cleared when Statistics OFF or when instrument is OFF or when test parameters are changed or when test is sa...

Page 80: ...Appendix B Parameters Thermo Electron Corporation B 4 ...

Page 81: ...ration A1 Aref f1 A2 Aref f2 Df optional f1 49 1 Protein concentration A2 Aref f3 A1 Aref f4 Df f2 3 48 f3 183 Ratio A1 Aref f4 75 8 A2 Aref dil vol 0 smp vol 1 DNA 260 280 Dilution Factor Df diluent vol sample volume Start wavelength µg mL with Scan sample volume 225nm Stop wavelength DNA concentration A1 Aref f1 A2 Aref f2 Df 325nm A1 260nm Protein concentration A2 Aref f3 A1 Aref f4 Df A2 280nm...

Page 82: ...order 595nm µg mL Std Conc of 0 20 40 60 80 100 Lowry standard Second order 550nm µg mL Std Conc of 0 100 200 500 1000 2000 Lowry micro Second order 750nm µg mL Std Conc of 0 20 50 100 200 500 Second order 562nm mg mL BCA standard Std Conc of 0 0 2 0 4 0 6 0 8 1 0 Second order 562nm µg mL BCA micro Std Conc of 0 0 5 1 2 5 10 Biuret First order through zero 540nm mg mL Std Conc of 0 2 4 6 8 10 Dire...

Page 83: ... A 8 400 x T coefficient units U 12 010 x G 7 050 x C M 1 cm 1 If entry does include U ε260 15 200 x A 9 900 x U 12 010 x G 7 050 x C Conversion Factor N A Molecular Weight x 103 µg mL Extinction Coefficient Calculation of Tm units A units T Tm 2 A T 4 G C C Oligos up to units G units C 20 bases in length Calculation of Tm units A units T Tm 81 5C 16 6log Na C DNA DNA hybrids units G units C 1 0 7...

Page 84: ...nits A units T Tm 78 C 16 6log Na C RNA RNA hybrids units G units C 1 0 7 Na 0 7 GC M molarity of cation 500 L P 0 35 formamide Fraction GC fraction of G and C form formamide in the sample L of base pairs P mismatching Conversion from µg mL and molecular pmol µL µg mL x 1000 pmol µL weight from Oligo calc DNA Mol Wt factor test ...

Page 85: ...ard N number of standards A A c where A absorbance c concentration A c is defined by an equation of the form A c a4c4 a3c3 a2c2 a1c a0 where a0 Y axis intercept a1 a4 coefficients The coefficients are computed using the least squares method A a1 c where A absorbance c concentration a1 slope The slope is calculated as a1 SXY SXX This model requires Slope is not equal to zero At least one standard d...

Page 86: ... specified domain and the message Curve cannot be used to determine unknown concentrations it may produce ambiguous results will appear when the curve is viewed where n degree of polynomial Note The calculation for the correlation coefficient only applies to first order linear regression curves first degree polynomials Absλ1 or Absλ1 Absref Absλ2 Absλ2 Absref Result Absλ1 factor 1 Absλ2 factor 2 o...

Page 87: ...C 7 BioMate 3 Operator s Manual Calculation Calculation s Graphs 3 Point Net 3 Point Net Absorbance sample curve Baseline corrected absorbance A2 A3 A1 A3 λ3 λ2 λ3 λ1 Appendix C Calculations ...

Page 88: ...Appendix C Calculations Thermo Electron Corporation C 8 ...

Page 89: ...on 1 1 Cell Correction program 1 4 3 3 3 4 3 5 3 8 3 9 3 12 3 15 3 16 3 17 3 18 3 20 3 24 3 25 3 26 Cell Growth 2 11 A 1 C 1 C 2 Cell holders cell holder accessories 3 4 5 1 5 2 6 3 Cleaning 6 1 6 2 Concentration measurements 3 6 3 10 Concentration units 3 1 3 2 3 6 3 7 Custom units 3 1 3 2 D Date and time setting 1 2 Direct UV 2 3 2 9 2 10 C 1 C 2 DNA 2 2 2 3 2 12 A 1 B 1 C 1 C 3 C 4 DNA with sca...

Page 90: ...ling 3 24 Photometric accuracy 4 1 4 3 4 4 Printer installing 5 5 3 Protein Measurements 2 1 2 7 R Replacement parts 6 3 C 2 RNA 2 2 2 3 2 5 2 12 C 1 S Safety i Sample compartment windows cleaning 6 2 Scanning 1 1 3 4 3 12 3 20 3 21 Screen contrast setting 1 2 Single Cell Platform 3 5 Single Position Cell Holder installing 5 Slope 3 12 3 13 C 5 SmartStart 3 2 Software password iii Specifications A...

Page 91: ...m 2 12 A 1 C 3 C 4 Total run time 3 17 B 3 U Unlock test 3 1 Utility parameters setting 1 1 1 3 W Warburg Christian 2 10 2 11 C 1 C 2 Warranty ii Wavelength accuracy 4 1 internal 4 1 standards 4 2 Z Z dimensions 1 1 1 2 I 3 BioMate 3 Operator s Manual Index ...

Page 92: ...Index Thermo Electron Corporation I 4 ...

Page 93: ......

Page 94: ...Item No 335904 10001 Rev G 06 03 Component No 335904 10063 Rev G 06 03 ...

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