boeco model s-30 life science analyser operating manual
Contents
1. 1100nm Pack 100 3 5ml plastic cuvettes 320 1100nm Pack 500 3 5ml plastic cuvettes 320 1100nm Dust cover Calibration set Xenon lamp module 10 x 10mm cell holder Replacement fuse 2A 28 S 30 REV A 02 05 SECTION 6 INTERFACING 6 1 SERIAL INTERFACE The Model S 30 has a bi directional RS232 interface set to 1200 baud 7 data bits odd parity 1 stop bit The 25 way D connector allows a standard one to one interconnection lead to be used as supplied with the 40 column printer A printout is initiated by pressing the PRINT key If the sample number is unity then the printout will include a header block The sample number is incremented every time the print key is pressed The following commands can also be sent to the Model S 30 via the serial interface ASCII D ord Sames as pressing the print key ASCII T lt CR gt Outputs transmission and wavelelength separated by an ASCII TAB character regardless of the Model S 30 operating mode For example 100 0 540 ASCII A lt CR gt Outputs absorbance and wavelength separated by an ASCII TAB character regardless of the Model S 30 operating mode For example 0 001 540 ASCII C lt CR gt Outputs concentration and wavelength separated by an ASCII TAB character regardless of the Model S 30 operating mode For example 123 4 540 ASCII V lt CR gt Outputs a voltage proportional to the monochromatic light level passing through the sample and wavelength separated by an ASCII TA2. EIT DILUTION FACTOR 50 GEBE E UM ITS Hom RESOLUT I Ory l WAVELENGTH 0260 ssDNA factor value EXIT DILUTION S200 0300 1 FACTOR 37 OGE G UNITE balm RESOLUTION 1 WAVELENGTH 268 RNA factor value EXIT DILUTION O150 6286 1 FACTOR 40 O0GE 0 UHITE pnl RESOLUTION l WAVELEHGTH zen S 30 REV A 02 05 EXIT DILUTION FACTOR UNITS RESOLUTION OLIGO factor value EXIT DILUTION G60 1 0000 1 FACTOR 360 000 0 UHITE Harml RESOLUT I Ory l WAVELENGTH zen Allows the user to exit this menu Allows the user to directly obtain the concentration of the original solution Enter the volume of the sample solution in ul Move the cursor to the right using the arrow key and enter the volume of the diluent in ul Use the enter key to store these values and return to the options listing To obtain an accurate result ensure the solution is thoroughly mixed prior to measurement Used for the concentration calculation where concentration Factor x Abs User adjustable calculation factor is displayed as an exponential value E 3 E 0 E 3 etc depending on the units of measure mantissa only is user adjustable Allows selection of the units used to display against the concentration reading The choice available ug ml ng ml mg ml ng ul Allows the user to specify the resolution of the displayed reading 1 0 1 0 01 0 001 The maximum resolution that concentration readings are displayed to can be set u
3. Resolution Accuracy Repeatability Bandwidth Factor Readout Outputs Light source Operating Temperature Maximum RH Input voltage Input power Size Weight 0 to 199 9 0 1 lt 0 5 340nm and 220nm 1 0 300 to 1 999A 0 001A lt 0 002A Hr after 30 minute warm up lt 0 001A 0A 400nm 300 to 9999 1 0 1 0 01 or 0 001 ppm mg l g l M 96 ug l ug ml mg ml ng ml or blank mode specific 198 to 1000nm inm 2nm 0 5nm 5nm typical 270nm 8nm over full wavelength range 0 to 199 9 1000 to 9999 Custom LCD Graphics Analogue 0 to 1 999V d c RS232 serial port Xenon flash lamp module 5 C to 40 C 80 115 230V a c 20 10 50 60Hz 50W 365 w x272 d x160 h mm 6kg 1 S 30 REV A 02 05 SECTION 2 INSTALLATION 2 1 UNPACKING Remove the Model S 30 from the packaging and ensure the following items are present 1 Model S 30 Life Science Analyser 2 Mains cable Pack 8 300yl disposable plastic cuvettes 035 132 4 Instruction manual 5 Optional accessories as ordered 2 2 INSTALLATION MAINS SUPPLY The Model S 30 is designed to operate on 115 230V a c supplies 20 10 50 60Hz The standard 2 metre mains cable supplied with the unit is fitted with an IEC type connector which can be plugged directly into the POWER IN socket on the rear panel The mains fuse is housed within the POWER IN socket When replacing the fuse the unit should be disconnected
4. a measurement Press any key to clear the above screen Select EXIT to clear the graphical screen Select EXIT again to return to the main measurement screen 14 S 30 REV A 02 05 Place a sample blank into the sample chamber and close the sample chamber lid Press the CAL key to initiate a calibration 0 l4 AL HBS 562 SETUP CURUE Remove the sample blank from the sample chamber and replace with the unknown sample The Absorbance value is read and plotted against the curve to determine the concentration value This value is presented in large figures at the top of the screen 151 0 180 562 SETUP CURUE 15 S 30 REV A 02 05 3 7 DIRECT UV This measurement mode differs from the other 4 in that it does not use a calibration curve for measurement but requires readings at 280 and 260nm and an answer calculated based on X 1 99 Ag X 0 76 Measurements can be made using 280nm only In this instance the Factor 1 value should remain as set 1 550 and the Factor 2 value should be set to 0 000 Select DIRECT UV from the Protein mode The following display will be shown palmi A 258 md READ SETUP Select the SETUP option EXIT FACTOR 1 1 556E 6 FACTOR 2 B FaGE 6 DILUTION mia8Beasmmnl UHI TS RESOLUTION l EXIT Allows user to exit this menu FACTOR 1 Value at which the Abs at 280nm gets multiplied by FACTOR 2 Value at which the Abs at 260nm gets multiplied by User adjustable calcu
5. following display will be shown S 0 b Q 057 7 114 0 260 SETIF It is recommended that setup parameters be reviewed prior to calibration or measurement to ensure the selected values are correct Select SETUP and the following options will be displayed EXIT FACTOR 1 UNITS PPP RESOLUTION LENS WAWELEHGTH 260 Allows the user to exit this menu FACTOR Used for the concentration calculation where concentration Factor x Abs UNITS Allows selection of the units used to display against the concentration reading The choice available 96 M g l mg l ppm none g dl RESOLUTION Allows the user to specify the resolution of the displayed reading 1 0 1 0 01 0 001 The maximum resolution that concentration readings are displayed to can be set up to 3 decimal places The instrument will automatically display concentration readings to the maximum possible resolution using this parameter WAVELENGTH Allows user setting of preferred wavelength for the test being performed Performing a measurement To perform a measurement it is necessary to set the unit to the required wavelength and perform a calibration Place a blank solution into the sample chamber and close the lid Press the CAL key The instrument will momentarily show CAL indicating that the calibration is being performed Once calibrated the display values will update to show ABS T and Concentration readings Remove the blank
6. general protein assay for batches of material for which yeild is not a problem Lowry assay The principal target is to reduce the high susceptibility to interference In comparison to the Biuret assay the sensitivity of this assay has greatly increased The Lowry method however is adversely affected by a wide range of non proteins Additives such as EDTA ammonia sulphate or Triton X 100 in particular are incompatible with the test 6 S 30 REV A 02 05 3 2 POWER ON SELF TEST Prior to switching the unit on check that the voltage select switch is set to the voltage supply being used When the unit is switched on a self test routine will automatically be performed POWER DH TESTS SYSTEM TEST PASS Ep DARK CAL PASS chamber iz Do not WAVELENGTH CAL door durins Bower on tests SYSTEM TEST This test checks the validity of the operating parameters The following messages may be dis played during this test CRITICAL ERROR CALIBRATION DATA FAILURE This error is non recoverable and means the calibration data for the detector is not working and therefore the unit cannot operate If this message is displayed during the test the manufacturer or local distributor should be contacted immediately for advice SYSTEM ERROR OPERATING PARAMETERS FAILURE This error is recoverable and means that the setup and operating parameters have been reset to their default values possibly due to memory corruptio
7. optimum performance with minimual maintenance It is only necessary to keep the external surfaces clean and free from dust The sample area should always be kept clean and any accidental spillage should be wiped away immediately To give added protection when not in use the unit should be disconnected from the mains supply and covered with the optional dust cover For longer term storage or re shipment it is recommended that the unit be returned to its original packing case NOTE The Model S 30 monochromator is a non serviceable unit and no attempt should be made to repair this item Failure to observe this recommendation will result in the loss of any Warranty Claim on this product In the unlikely event of the monochromator requiring service or calibration it is essential that the manufacturer or your local distributor be contacted immediately for advice 4 2 LIGHT SOURCE REPLACEMENT The only routine maintenance which may be required is the replacement of the light source if this fails Failure should be suspected if the instrument fails wavelength calibration during initialisation or the instrument is unable tocalibrate to zero absorbance when the CAL key is pressed The message LIGHT LEVEL TOO LOW will be displayed The xenon flash lamp module is available from the manufacturer or your local distributor refer Section 5 2 Spares CAUTION The following safety precautions should be observed prior to attempting the light source replaceme
8. side of a user entered centre wavelength 250 950nm at 1nm steps Select the PURITY SCAN MENU Ed SETUF The following display will then be shown Remember to Perform blank calibration Ead SETUF CAH Select SETUP and adjust the wavelength value as appropriate for the test being performed EXIT WAUELEHWGTH 21 S 30 REV A 02 05 When the required wavelength value has been entered and confirmed place a sample blank into the sample chamber and close the lid Press CAL and the instrument will perform a calibration as shown Ead SETUF CAH Remove the sample blank from the sample chamber and insert the unknown sample Close the sample chamber lid Select SCAN The instrument will perform the scan as shown ERIT SETUF ERIT SETUF The display will then update to show peak absorbance and the wavelength of the peak Feak barnn S91 Abs anm ERIT 22 S 30 REV 02 05 dsDNA ssDNA RNA and OLIGO MODES NOTE These 4 modes although using different factor values are all performed using the same procedures as detailed below Measurement method is Photometrics Absorbance T and Concentration values are displayed simultaneously i e they are not individually selectable Select the appropriate measurement mode from the main menu and the following display will be FES shown Select SETUP 1 260 dsDNA factor value
9. the following methods of analysis B C A second order curve fit quadratic 562nm Bradford second order curve fit quadratic 590 or 595nm Lowry second order curve fit quadratic 550 750nm or 500 750nm Biuret first order curve fit quadratic 540 or 550nm Direct UV multi wavelength 3 S 30 REV A 02 05 B C A Bicinchoninine acid assay This test is a highly regarded alternative to the Lowry assay being much easier to carry out and sensitivity can be varied using different temperatures The dye complex is very stable This test however can be susceptible to interference but its insensitivity to detergents is comparable to the Lowry method Bradford assay This method is twice as sensitive as the B C A or Lowry test and is the most sensitive quantitative dye assay It is the easiest to handle and the most rapid method It also has the additional advantage that a series of reducing substances e g DTT and mercaptoethanol have no adverse effect on results It is however sensitive to detergents The main disadvantage with this method is that identical amounts of different standard proteins can cause considerable differences in the resulting absorption coefficients Biuret assay The principle of the Biuret is similar to that of the Lowry However it involves a single incubation of 20 minutes There are very few interfering agents ammonium salts being one such agent The Biuret consumes more material This assay is a good
10. the highlight around menu screen options If editing a numeric value the highlighted digit can be altered If the highlight is moved off the left most digit the data editing will be aborted and the previous value will be re instated used to select the highlighted menu option or to store the current parameter being entered initiates a calibration routine absorbance zero provides a printout of the current reading with an incremental sample number When pressed for the first time after a calibration the print out will give calibration information The incremental sample number will be reset after a calibration 3 S 30 REV A 02 05 2 4 INPUTS OUTPUTS DANGER udgornect paver before TAINS removing covers SZ 1 ROCKER SWITCH 2 POWER IN SOCKET 3 OUTPUT SOCKETS 4 OUTPUT SOCKET On off switch for the unit IEC type connection socket for mains cable Analogue output Output socket for 25 way for RS232 S 30 REV A 02 05 SECTION 3 OPERATION 3 1 GENERAL PRINCIPLES The Model S 30 is a grating spectrophotometer with a selectable wavelength range of 198 1000nm For specific nucleotide and protein assays software is on board to facilitate rapid analysis of these compounds Using spectroscopy the quantitative analysis of these assays has now become a routine method in many laboratories It includes absorption measurement primarily in the ultraviolet range Proteins are measured directly at 280nm nucleic acids at 260
11. 5 CTS To PC DSR6 M 6 DSR To PC DCD 8 8 DCD To PC DTR 20 20 DTR From PC GND 7 7 GND MODEL S 30 IBM PC XT 9 way D TXD 2 3 TXD From PC RXD 3 2 RXD To PC 4 7 RTS From PC CTS 5 8CTS To PO DSR6 6DSR PC DCD 8 Y 1 DCD To PC DTR20 4 From GND 7 5 GND NOTE The interface cable kit order code 542 009 can be used to implement the above connections 6 3 RECORDER OUTPUT This is available via the 4mm rear panel sockets The level is proportional to the displayed concen tration reading Concentration 1mV per concentration unit 30 S 30 REV A 02 05 EC Declaration of Conformity The BOECO Model S 30 complies with the following European Standards EN 61326 1998 Electromagnetic compatibility Electrical equipment for measurement control and laboratory use EN 61010 1 2001 Safety requirements for electrical equipment for measurement control and laboratory use Following the provision of EMC Directive 89 336 EEC and Low Voltage Directive 73 23 EEC Carl Warren Technical Manager Jenway Gransmore Green Felsted Dunmow Essex CM6 3LB England 31 S 30 REV A 02 05 Method PROTEIN Lowry BCA Bradford Biuret
12. B character For example 1234 5 540 ASCII Z lt CR gt Calibrates a zero absorbance if the xenon lamp is on SO CR command or zero transmittance if the lamp is off SGC lt CR gt command ASCII SC lt CR gt Switches the xenon lamp off This allows 0 transmittance to be calibrated ASCII SO lt CR gt Switches the xenon lamp on This allows 100 transmittance zero absorbance to be calibrated The lamp must be on for normal measurements ASCII Gnnn lt CR gt Commands the Model S 30 to go to the wavelength nnm For example G540 CR will set the wavelength to 540nm ASCII Fxxxx x lt CR gt Sets the concentration factor to xxxx x For example F1000 CR will set the factor to 1000 NOTE lt CR gt is an ASCII carriage return character The commands provide an output which can readily be incorporated into most spreadsheet software packages 29 S 30 REV A 02 05 6 2 RS232 OUTPUT The bi directional RS232 interface is availble on the rear panel 25 way D connector The connections are as follows TXD 2 INPUT TO MODEL S 30 RXD 3 OUTPUT FROM MODEL S 30 RTS 4 LINKED TO CTS CTS 5 LINKED TO RTS DSR6 OUTPUT FROM MODEL S 30 DCD 8 OUTPUT FROM MODEL S 30 DTR 20 INPUT TO MODEL S 30 must be active Suggested interconnections are detailed below MODEL S 30 IBM PC XT 25 way D TXD 2 2 TXD From PC RXD 3 ___ 3 RXD To PC RTS 4 ARTS From PC CTS5
13. BOECO MODEL S 30 LIFE SCIENCE ANALYSER OPERATING MANUAL 636 315 REV A 02 05 Safety This is important information please read carefully before installing or using this instrument 1 The Model S 30 life science analyser is designed for operation by trained personnel that are aware of the principles and applications involved For further help and advice please contact your local distributor e mail info boeco com or visit www boeco com The Model S 30 life science analyser is a sensitive electronic and optical instrument designed for use in a laboratory environment Careful adherence to the installation instructions must be observed If in doubt contact a relevant and competent authority for advice before proceeding In addition to observing the instructions detailed in the Operating Manual and Service Manual for this instrument all installation operating and service personnel must be aware of and employ a safe system of work Voltage levels hazardous to life are present in this instrument for personal safety only trained engineers aware of the risk and avoidance of electric shock should remove protective covers from the instrument This instrument is designed for minimal maintenance which must be carried out carefully following the procedures detailed in this manual All safety instructions in these procedures as well as those defined locally for the area or environment where the work is being carried out must be observed Other t
14. DNA MEASUREMENT DNA Purity Absorbance difference 260 280 Absorbance difference 260 230 Direct UV DNA MEASUREMENT References References Lowry O H Rosebrough N J Farr A L and Randall R J 1951 J Biol Chem 193 265 275 Smith P K Krohn R L Hermanson G T Mallia A K Gartner F H Provenzano M D Fujimoto E K Goeke N M Olson B J and Klenk D C 1985 Anal Biochem 150 76 85 Bradford M 1976 Anal Biochem 72 248 254 Ohnishi S T Barr J K Anal Biochem 86 193 1978 Layne E 1951 Methods in Enzymology 3 447 454 Groves W E Davis F C and Sells B H 1968 Anal Biochem 22 195 209 Warburg O Christian W 1941 Biochem Z 310 384 Kalb V F Bernlohr R W 1977 Anal Biochem 82 362 371 Maniatis T Fritch E F Sambrook J 1989 Molecular Cloning A Laboratory Handbook 3 32 S 30 REV A 02 05
15. e data editing will be aborted and the previous value will be re instated used to select the highlighted menu option or to store the current parameter being entered initiates a calibration routine provides a printout of the current reading with an incremental sample number When pressed for the first time after a calibration the print out will give calibration information The incremental sample number will be reset after a calibration 3 4 GLOBAL SETUP PARAMETERS INSTRUMENT SETUP MENU This option will allow all instrument related setup i e non mode specific parameters to be set up The options available through the Instrument Setup menu are as follows EXIT TIME DATE DATEFORMAT HELP TEXT CELL EXIT TIME aa aa DATE Od sod DATE FORMAT OD HELF TERT YES CELL CUUETTE Returns to the main menu screen Allows the user to set the current time into the instrument in the form of HH MM SS Allows the user to set the current date into the instrument in the form of either DD MM YY or MM DD YY depending on the setting of the date format field Allows the user to choose between either the DD MM YY formatting of dates or MM DD YY formatting The selected format is applicable to all displayed dates Additional help messages will be displayed if the YES option is selected These messages provide assistance with general operation If the message does not appear a second time it is because it has already b
16. een shown since power on Allows choice of capillary or cuvette On selection this will automatically correct the ABS value for the appropriate path length to its displayed value per cm 8 S 30 REV A 02 05 SECURITY EXIT INSTRUMENT LOCK SECURITY CODE EXIT IHSTRUMENT LOCK Ho SECURITY CODE BBL Returns to the main menu screen Yes or no option When set to Yes the up and down arrow keys are disabled within set up screens thus preventing operating parameters of the unit from being changed Standard curve and absorbance zero calibrations are possible when the instrument lock is activated The Photometrics mode is not affected by the instrument lock feature When the instrument lock is active entry of a locking code 0 to 999 set when the instrument lock is activated is required to re enter the security option PHOTOMETRICS CHA RHA MODE PROTEIN MODE IHSTRUMEHT SETUP SECURITY Needed to get back into security menu to turn it off 0 999 If no security number is available turn the unit off and then on holding down the enter key This then resets to no security code and also any other previously set parameters Will prompt first message refering to operating parameters indicated by a failure in System Test 9 S 30 REV A 02 05 3 5 PHOTOMETRICS MODE NOTE Absorbance T and Concentration values are displayed simultaneously i e they are not individually selectable Having selected PHOTOMETRICS from the main menu the
17. from the mains supply In the event of the fuse failing after replacement it is advisable to consult with the manufacturer or your local dealer before proceeding further Fuse rating 2A F fast blow type NOTE The unit should be positioned within 1 5 metres of an earthed mains supply VOLTAGE SELECT NOTE When changing the voltage select switch position always ensure the fuse rating is correct Before attempting to change the voltage select disconnect the unit from the mains supply with draw the fuse holder form the power input socket and remove the fuse Extract the grey fuse retainer and rotate so that the correct voltage is visible through the aperture in the fuse holder Replace the fuse retainer in its holder fit the correct fuse and push assembly back into the power input socket MAINS CONNECTIONS A suitable plug should be connected to the 3 wires on the mains lead These are colour coded to conform with the internationally recognised standard such that BROWN LIVE BLUE NEUTRAL GREEN YELLOW EARTH IMPORTANT THE UNIT MUST BE EARTHED The Green Yellow wire in the a c supply cable must be connected to a properly grounded terminal 2 S 30 REV A 02 05 2 3 CONTROLS 1 UP DOWN KEYS 2 LEFT RIGHT KEYS 3 ENTERKEY 4 CAL KEY 5 PRINTKEY used to move the highlight around menu screen options unless editing a parameter In this instance these keys are used to adjust the highlighted parameter used to move
18. han for those items defined in the maintenance procedures herein there are no user serviceable items in this instrument Removal of covers and attempted adjustment or service by unqualified personnel will invalidate any warranty and incur additional charges for repair Reference should always be made to the Health and Safety Data for any chemicals or reagents used All available information advice and warnings on the handling storage use and disposal of such must be carefully observed When not available this data must be requested from the supplier before proceeding in any way It is important that good laboratory practice is observed when handling samples chemicals reagents and ancillary equipment in order to carry out measurement and analysis with this instrument Suitable safety and personal protective equipment must be used at all times If it is suspected that safety protection has been impaired in any way the spectrophotometer must be made inoperative and secured against any intended operation The fault condition must be reported to the appropriate servicing authority In all such reports the model number and serial number of the spectrophotometer must be quoted S 30 REV A 02 05 CONTENTS SECTION 1 SECTION 2 SECTION 3 SECTION 4 SECTION 5 SECTION 6 BOECO S 30 LIFE SCIENCE ANALYSER OPERATING MANUAL INTRODUCTION Instrument Description Instrument Specifications INSTALLATION Unpacking Installation Controls I
19. he instrument display will update to show the following EMIT WE Place std 1 in the sample chamber and Fress CAL A 2 d Tua Tu Du and then update requesting the next standard to be placed in the sample chamber until the specified number of standards has been calibrated EMIT VIEW Place std in the sample chamber and Press CAL If it is necessary to abort the calibration sequence before all standards have been measured this can be carried out by pressing any keys other than CAL or ENTER Only information entered up to that point will be retained 13 S 30 REV A 02 05 Once entry of these values is completed selection of the VIEW option will allow the user access to the graphical calibration curve constructed from these values An X displayed on the graph indicates the position of a calibration point STATISTICS Selecting STATISTICS from this menu will show the terms used for the calculated quadratic curve fit The curve fit is of the form y ax bx c so the statistics page shows the ax bx and c terms separately Pressing any key again allows the user to view the x and y min max values i e the axis limits to which the graph is plotted The x axis concentration the y axis absorbance If unlocked it is possible to adjust amend the standard or Absorbance values by using the right arrow key to move over to Absorbance values Performing
20. lation factors are displayed as exponential values E 3 E 0 E 3 etc depending on the units of measure mantissa only is user adjustable DILUTION Allows the user to directly obtain the concentration of the original solution Enter the volume of the sample solution in ul Move the cursor to the right using the arrow key and enter the volume of the diluent in ul Use the enter key to store these values and return to the options listing To obtain an accurate result ensure the solution is thoroughly mixed prior to measurement UNITS Allows the user to select the preferred measurement unit ug ml mg ml ng ml ng ul g dl mg dl RESOLUTION Allows the user to select the preferred resolution 1 0 1 0 01 or 0 001 The final figure shown will be reduced in resolution if it not possible to show all the required decimal places on the screen If the READ option is selected prior to performing a calibration the following message will be displayed balm Ferform CAL First please ERIT SETIF S 30 REV A 02 05 When all set up parameters have been entered exit this option Place a sample blank into the sample chamber and close the lid Press CAL and the instrument will calibrate at 280 and 260nm even when only 280nm is required ERIT SETUF EMIT The display will update to show zero at both points 268 268 GG8Ab s 4 BEERS ERIT SETUP Remove the sample blank from the chamber and insert the u
21. lume of the sample solution in ul Move the cursor to the right using the arrow key and enter the volume of the diluent in ul Use the enter key to store these values and return to the options listing To obtain an accurate result ensure the solution is thoroughly mixed prior to measurement CORRECTION YES NO option A reference wavelength is optional If this is to be used in calculations then YES should be selected WAVELENGTH If the correction option is set to yes then the wavelength must be specified UNITS Allows the user to select the preferred measurement unit ug ml mg ml ng ml ng ul RESOLUTION Allows the user to select the preferred resolution 1 0 1 0 01 or 0 001 The final figure shown will be reduced in resolution if it not possible to show all the required decimal places on the screen 18 S 30 REV A 02 05 When all set up parameters have been entered exit this option If the READ option is selected prior to performing a calibration the following message will be displayed Ferform CAL First please ERIT SETIF Place a sample blank into the sample chamber and close the lid Press CAL and the instrument will calibrate at 260 280nm or 260 230nm depending on the mode selected CAL 26Bnm CAL 288nm RATIO RATIO ERIT SETUP ERIT SETUP If the reference wavelength option is selected a third calibration will be performed at the nominal reference wavelength of 320nm All 3 values will be shown on the instrumen
22. n To continue press any key DARK CALIBRATION TEST The message SYSTEM ERROR DARK LEVEL CALIBRATION FAILURE may be displayed if the unit receives too much light when trying to perform a dark level calibration Pressing any key clears this message and the unit will retry calibration It will repeat this process until it passes the test satisfactorily WAVELENGTH CALIBRATION This part of the self test checks the optical alignment of the unit If it fails this test an error message SYSTEM ERROR WAVELENGTH CALIBRATION FAIL URE will be displayed This message is displayed permanently on screen and can only be re moved by powering the unit off If this message is displayed during the test the manufacturer or local distributor should be contacted immediately for advice Once the power on self test is successfully completed the main menu will be displayed PHOTOMETRICS DHAYRHA MODE PROTEIN MODE IHSTRIUMEHT SETUP SECURITY S 30 REV A 02 05 3 3 KEYPAD OPERATION The following information applies to all setup and operating modes UP DOWN KEYS used to move the highlight around menu screen options unless editing a parameter In this instance these keys are used to adjust the highlighted parameter LEFT RIGHT KEYS used to move the highlight around menu screen options If editing a ENTER KEY CAL KEY PRINT KEY numeric value the highlighted digit can be altered If the highlight is moved off the left most digit th
23. nknown sample Close the sample chamber lid Select READ and the instrument will measure at both points even when only 280nm is required MEASURE 8 z8B8nm MEASURE 8 66m ERIT SETUF ERIT SETUP Once the measurement sequence has been successfully performed the display will update to show the final sample reading 0 2017 268 h 26H B lirzHbs 8 826Abs ERIT SETUP 17 S 30 REV A 02 05 3 8 DNA RNA MODE Select the DNA RNA MODE option from the main menu Esty 260250 PURITY dsDHH ssDNA RHH OLIGO WARIAELE RATIO Set up and measurement procedures are the same for 260 280nm and 260 230nm modes as detailed below Select the appropriate mode of operation 260 280nm or 260 230nm and the following screen will be displayed Pas 280 FATIO d SETUF Select SETUP and the following menu will be displayed ERIT FACTOR 1 Ez 9SRE 8 FACTOR 2 26 HBBBETEB DILUTION S826 8856 1 CORRECT OM HO WAVELENGTH 328 UHITE Holm RESOLUT I OH 1 Allows the user to exit this menu FACTOR 1 Value at which the Abs at 260nm gets multiplied by FACTOR 2 Value at which the Abs at either 230 or 280nm gets multiplied by User adjustable calculation factors are displayed as exponential values E 3 E 0 E 3 etc depending on the units of measure mantissa only is user adjustable DILUTION Allows the user to directly obtain the concentration of the original solution Enter the vo
24. nm and colorimetric protein determination is carried out within the range of 550 to 600nm Nucleic Acid Determination DNA RNA and oligonucleotides can be measured directly in aqueous solutions in a diluted or undiluted form Aqueous buffers with low ion concentrations e g TE buffer are ideal for this method The concentration is determined by measuring at 260nm against a blank and then evaluating against factor The absorption of 1 OD A is equivalent to approximately 50ug ml dsDNA 37ug ml ssDNA 40ug ml RNA or 30ug ml for oligonucleotides Purity determination of DNA interference by contaminants can be recognised by the calculation of ratio The ratio A260 A280 is used to estimate the purity of nucleic acid since proteins absorb at 280nm Pure DNA should have a ratio of approximately 1 8 pure RNA 2 0 Absorption at 230nm reflects contamination of the sample by substances such as peptides phenols aromatic compounds or carbohydrates In pure samples the ratio should be approximately 2 2 Referring to a blank value where no absorption should occur is commonly required On the S 30 this default reference is 320nm Should you then wish to change or modify these wavelengths this flexibility is in built Protein Determination Several analytical procedures can be used to determine the protein content of a preparation Evaluation can be carried out either via a calibration curve or a factor using up to 6 standards The Model S 30 uses
25. nputs Outputs OPERATION General Principles Power On Self Test Keypad Operation Global Setup Parameters Photometrics Mode Protein Mode Direct UV DNA RNA Mode Good Practice Guidelines MAINTENANCE General Light Source Replacement OPTIONAL ACCESSORIES Optional Accessories Spares INTERFACING Serial Interface RS232 Output Recorder Output EC Declaration of Conformity 3 1 3 2 3 3 3 4 3 5 3 6 3 7 3 8 3 9 4 1 ooo On S 30 REV A 02 05 SECTION 1 INTRODUCTION 1 1 INSTRUMENT DESCRIPTION The Model S 30 is optimised for use by the life science research chemist for the routine purity tesing of proteins DNA and bacterial samples Fully menu driven operation allows ease of use without the need for detailed knowledge of spectroscopy and makes the product ideal for use in teaching laboratories Features include dedicated operating modes for DNA RNA oligonucleotide and protein analysis Includes DNA programs auto ratio and direct calculation of ssDNA dsDNA RNA and oligonucleotide concentrations and protein calculations including Lowry Bradford BCA and direct reading with display of calibration curve Scanning capability for peak purity check 1 2 INSTRUMENT SPECIFICATIONS Transmittance Range Resolution Stray light Photometric accuracy Absorbance Range Resolution Photometric stability Photometric noise Concentration Range Resolution Units Wavelength Range
26. nt procedure 1 DISCONNECT THE UNIT FROM THE MAINS SUPPLY PRIOR TO REMOVING THE TOP COVER 2 SAFETY GLASSES MUST BE WORN WHEN UV EMISSIONS ARE PRESENT 3 DO NOT TOUCH THE LIGHT OUTPUT MODULE OF THE LAMP MODULE 26 S 30 REV A 02 05 Remove the 4 case screws as shown Lift the top cover clear taking care not to strain the ribbon cable Remove the 2 xenon lamp retaining thumb screws Remove the connector from the rear of the xenon lamp When replacing the lamp ensure it is pushed fully up into the optics housing before tightening the thumb screws Refit the top cover taking care with the ribbon cable Refit the 4 case screws 27 S 30 REV A 02 05 SECTION 5 OPTIONAL ACCESSORIES 5 1 OPTIONAL ACCESSORIES The following list of items is available for use with the Model S 30 BOE 035 124 BOE 035 132 BOE 632 001 BOE 632 031 BOE 634 001 BOE 021 041 BOE 630 005 BOE 035 079 BOE 035 087 BOE 035 029 BOE 035 086 BOE 035 027 BOE 060 087 BOE 060 084 BOE 060 229 BOE 630 028 BOE 035 088 5 2 SPARES BOE 012 094 BOE 630 004 BOE 016 021 UV quartz ultra micro 50ul cuvette Pack 100 500ul UV plastic cuvettes 220 900nm Sipper pump 230V version Sipper pump 115V version 4 position cell holder DC AC power converter 10 100mm cell holder 100 x 10mm glass cell 50 x 10mm glass cell 40 x 10mm glass cell 20 x 10mm glass cell 10 x 10mm glass cell Pack 100 1ml plastic semi micro cuvettes 320
27. osilicate glass The use of soda glass should be avoided where possible as leaching can occur during prolonged contact giving erroneous results Glass cuvettes should be thoroughly cleaned after use Discard when scratches become evident in polished surfaces Chemical reagents should wherever possible be of high grade quality Contamination can cause problems even at very low levels Diluents i e water or solvents must be free from impurities There are some substances which do not follow Beer s Law When attempting a new method it is advised that linearity checks should be performed over the range of concentrations being used This can be carried out by preparing a quantity of known strength solutions and checking the results a Deviations from Beer s Law may occur at high concentrations by association of molecular ionic species b Deviations from Beer s Law may occur at low concentrations by variation in hydration introducing changes in the nature of complex ions c Absorption which does not obey Beer s Law will require a graph of known standards to be plotted This should indicate Reading vs Concentration The reading obtained from the unknowns can then be related to the concentrations from the graph Samples and standards can outgas when left in the cuvette Bubbles formed on the cuvette walls will cause reading errors 25 S 30 REV A 02 05 SECTION 4 MAINTENANCE 4 1 GENERAL The Model S 30 is designed to give
28. p to 3 decimal places The instrument will automatically display concentration readings to the maximum possible resolution using this parameter WAVELENGTH Allows user setting of preferred wavelength for the test being performed Performing a measurement To perform a measurement it is necessary to calibrate the unit first Place a blank solution into the sample chamber and close the lid Press the CAL key The instrument will momentarily show CAL indicating that the calibration is being performed Once calibrated the display values will update to show ABS T and Concentration readings Remove the blank solution from the sample chamber The instrument is now ready to perform a measurement The instrument will now continually perform a live measurement The sample value will be shown directly as ABS T and Concentration purml 0 081 7 83 0 260 SETUP 24 S 30 REV A 02 05 3 9 GOOD PRACTICE GUIDELINES 1 To ensure accurate results are obtained the sample area lid should be kept in the closed position during measurement The styrene cuvettes supplied with the unit are disposable i e ideally they should be used once and then thrown away Some repeat use is possible providing extreme care is taken during cleaning to ensure no damage occurs to the polished surface Plastic cuvettes are not suitable for use with organic solvents Glassware used in the preparation of standards should be of high grade bor
29. solution from the sample chamber The instrument is now ready to perform a measurement The instrument will now continually per form a live measurement The sample value will be shown directly as ABS T and Concentration 10 S 30 REV A 02 05 3 6 PROTEIN MODE Select PROTEIN MODE from the main menu options The following display will be shown E C H BRADFORD LOWE EIURET DIRECT Lu NOTE B C A Bradford Lowry and Biuret although using different wavelength settings are all performed using the same procedures as detailed below Measurement method is by Standard Curve i e absorbance versus concentration Curve fit Quadratic Select SETUP and the following display will be shown EXIT WAWELEHWGTH D67 UHITS m3 41 RESOLUTION Allows the user to exit this menu WAVELENGTH Allows the user to select the appropriate wavelength if different from the standard default values of B C A 562nm Bradford 590nm Lowry 750nm Biuret 540nm UNITS Allows the user to select the preferred measurement unit ug ml mg ml ng ml none g dl mg dl RESOLUTION Allows the user to select the preferred resolution 1 0 1 0 01 or 0 001 The final figure shown will be reduced in resolution if it not possible to show all the required decimal places on the screen 11 S 30 REV 02 05 Prior to sample measurement it is necessary to construct a curve using a number of standards Select CURVE and the following display
30. t display Pas CAL 328nnm 26H 290 GG0Ab s 8 88 Abs FATIO A FAT IO ABBAS ERIT SETUF ERIT SETUP The DNA calculation will be performed by reading the absorbances at the required wavelengths and calculating the answer based on the pre entered factors 19 S 30 REV A 02 05 Remove the sample blank from the chamber and insert the unknown sample Close the sample chamber lid Select READ and the instrument will measure at 2 or 3 points depending on selec tion non selection of reference wavelength option MEASURE 26B8nm MEASURE z8Bnm MEASURE 3eBnm A 320 RATIO A 320 RATIO A xz RATIO EXIT SETUF EXIT SETUF EXIT SETUF Once the measurement sequence has been successfully performed the display will update to show the final sample reading 0 5047 26 290 B l zhbz 85 194Bbsz A F20 RATIO B B34Hbes 6 557 d SETUF VARIABLE RATIO This mode operates in the same way as the 260 280nm and 260 230nm modes with the additional benefit of the ability to specify wavelength values where peaks are not at 260 280 or 260 230 This enables fine adjustment for greater accuracy ET FACTOR 1 49 188E 6 FACTOR 2 2 408E 8 DILUTION aazcecazesl WAWELENGTH 1 260 WAWELEMGTH 2 CORRECTION WAVELENGTH UHITS RESOLUTION EXIT Allows the user to exit this menu FACTOR 1 Value at which the Abs at Wavelength 1 gets multiplied by FACTOR2 Value at which the Abs at Waveleng
31. th 2 gets multiplied by User adjustable calculation factors are displayed as exponential values E 3 E 0 E 3 etc depending on the units of measure mantissa only is user adjustable DILUTION Allows the user to directly obtain the concentration of the original solution Enterthe volume of the sample solution in ul Move the cursor to the right using the arrow key and enter the volume of the diluent in ul Use the enter key to store these values and return to the options listing To obtain an accurate result ensure the solution is thoroughly mixed prior to measurement WAVELENGTH 1 Allows adjustment of the first wavelength value 260 WAVELENGTH 2 Allows adjustment of the second wavelength value 230 CORRECTION YES NO option A reference wavelength is optional If this is to be used in calculations then YES should be selected 20 S 30 REV A 02 05 WAVELENGTH If the correction option is set to yes then the wavelength must be specified UNITS Allows the user to select the preferred measurement unit ug ml mg ml ng ml ng ul g dl mg dl RESOLUTION Allows the user to select the preferred resolution 1 0 1 0 01 or 0 001 The final figure shown will be reduced in resolution if it not possible to show all the required decimal places on the screen PURITY SCAN This mode provides a graphical representation of the Abs range and determines the peak absorbance It allows a sample to be scanned absorbance versus wavelength 50nm either
32. will be shown EMIT VIEW EMIT WIE METHOD OF STOS Curve can be calibrated by Pressing the EXIT Allows the user to exit this menu VIEW Allows the user to view the calibration curve as a graphic METHOD Allows the user to select which of the 10 methods available they are working on numbered 0 9 When the method number is changed the data previously displayed is stored against the previous number and you are presented with the data stored against thecurrent method number NO OFSTDS Number of standards being used for the calibration curve NO ppm ABS Column headings for the calibration points NOTE Date and time will be shown at the bottom of the display This indicates the last time the method was modified Measurements will be displayed in Absorbance only Enter the values of the standards being used with the lowest concentration value being entered first lowest to highest value in the centre column having first selected the unit of measurement required A minimum of 3 standards to a maximum of 6 is required Standard curve absorbance values can be manually entered if known by the user or by calibration with known standard solutions Press the CAL key and the following message will be displayed EMIT MTELN Press CAL kea La zero abzorbance Samele blank Press the CAL key again with a sample blank present in the sample chamber The display will momentarily show CAL S 30 REV A 02 05 and t
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