SimpliNano™ - GE Healthcare Life Sciences
Contents
1. ACCESSORIES INSTALLATION 7 1 Printer installation The user can install a printer Step 1 Remove the power cable and turn the instrument over onto a soft surface taking care not to damage the sampling head Release the outermost screws using the Allen key provided Step 2 Turn the instrument back over and remove the accessory covers Step 3 Attach the printer cable 29088793 AB 05 2014 25 29088793 AB 05 2014 26 Step 4 Lower the printer onto the locating bosses Step 5 Replace the top cover plate invert the instrument and replace the cap head screws at locations A and B shown in step 1 Step 6 Switch the instrument on and go to utilities instrument preferences and select the Built in printer 7 2 After sales support For all technical support and advice on SimpliNano please contact your supplier GE Healthcare s website is http www gelifesciences com or http www gelifesciences com contact Support agreements that help you to fulfil the demands of regulatory guidelines concerning GLP GMP are available e Calibration certification e Certificated engineers and calibrated test equipment e Approved to ISO 9001 standard Choice of agreement apart from break down coverage can include e Preventative maintenance e Certification Accessories Built in Printer accessory 28 9182 27 Spare paper for printer 20 rolls 28 9182 26 29088793 AB 05 2014 ef 8 IROUBLESHOOTI2. Toggle Graph on off The graph shows a wavescan plot across the range 220 nm to 320 nm with cursors denoting 230 260 280 and if Background Correction is selected 320 nm Sample Number add a prefix to the sample number and reset the EZ Sample Number incrementing number to the desired value Save Method Save Method use the alpha numeric keys to enter a name for the method Auto Print and press Save e Auto Print toggles Auto Print on off Exit options by pressing Esc or wait 29088793 AB 05 2014 18 4 5 Absorbance Concentration The procedure is as follows Step 1 Press 4 to select Absorbance Concentration mode Step 2 Select the wavelength using either arrow or alphanumeric keys Press the down arrow Step 3 Enter the measurement mode Absorbance Concentration using factor or single standard If Absorbance selected press OK to enter the Results screen If Factor or Standard selected press the down arrow Step 4 if factor selected Enter relevant factor and units Press OK Pu to enter the Results screen Step 5 If Standard selected Enter the concentration and units of the standard to be measured using the keypad numbers Press OK to enter the Results screen Step 6 Pipette the reference sample Press key This reference will be used for alll subsequent samples until changed If QA is switched on the sample will need to be replaced and the key pressed again Step 7 Pipette the sa
3. ok W Cancel Options select using keypad numbers Return to DNA Parameters screen step 1 above Q Print result via selected method Toggle Graph on off The graph shows a wavescan plot across the range 220 nm to 320 nm with cursors denoting 230 260 280 and if Background Correction selected 320 nm Sample Number add a prefix to the sample number and reset the incrementing number to the desired value Q Save Method use the alpha numeric keys to enter a name for the method and press Save Auto Print toggles Auto Print on off Exit Options by pressing Esc or wait 4 5 RNA measurement The procedure is as follows Step 1 Press 2 to select RNA Parameters mode Step 2 dilution factor known Enter the dilution factor using the keypad numbers Range 1 00 to 9999 Use the C button to backspace and clear the last digit entered OR Step 2 calculate dilution factor Press to enter the Dilution Factor screen see second image to the right Enter the volume of the sample using the keypad numbers range 0 01 to 9999 Press the down arrow Enter the volume of the diluent using the keypad numbers range 0 01 to 9999 Press OK to calculate the dilution factor and return to the Parameters screen OR press co to cancel the selections and return to the Parameters screen Step 3 Select whether the background correction at 320 nm is used or not with the left
4. Press the down arrow Enter the Month Press the down arrow Enter the Year Press the down arrow Enter the Hour Press the down arrow Enter the Minute Seconds are zeroed when OK is pressed Press OK to store the settings and return to the Utilities screen OR Press Cancel Esc to return to the Utilities screen without storing the time 6 2 Regional Sets Language and Number Format The procedure is as follows Select a Language Options are English French German Spanish Italian Chinese or Japanese Press the down arrow Set the decimal point style Options are or Press OK to store the settings and return to the Utilities screen OR Press Cancel Esc to return to the Utilities screen without storing the settings o Cancel 29088793 AB 05 2014 23 6 5 Printer Sets up printing options The procedure is as follows Select whether Auto Print is on or off using the left and right arrows When Auto Print is on the results are automatically printed after a measurement is taken When it is off printing has to be initiated manually This can also be set using the Options key in each application or method The default is Off Press the down arrow Select how the data are sent Options are Built in internal printer or to a computer via USB port Press OK to store the settings and return to the Utilities screen OR Press Cancel Esc to return to the Utilities
5. and right arrows Press the down arrow Step 4 Select the Units of measurement using the left and right arrows Options ug ml ng ul ug ul Press the down arrow Step 5 Enter the factor using the keypad numbers The default value is 40 the range is 0 01 to 9999 Step 6 Press OK to enter the Results screen and start taking measurements OR ec to return to the Home page 29088793 AB 05 2014 16 5 3 Print ox Parameters QR Sample Number Save Method amp uto Print v RANA Parameters Dilution Factor Factor Background a OK EB Cancel Results Screen Step 7 A230 oosa Sample Pipette the reference sample Press Key This reference will be used for all 200 0250A A280 0 166 subsequent samples until changed If QA is switched on the sample will need to be sll Concentration replaced and the key pressed again l 10 3 Ln Step 8 L Mns b Clean the sample port and pipette sample Press 4 to measure at the selected 1 news DES s wavelengths and display results 230 saa Repeat step 8 for all samples Press co to return to the Home page OR press to display the following options Options select using keypad numbers ap Parameters Return to Parameters screen step 1 above Pin Q Print result via selected method Graph Toggle Graph on off The graph shows a wavescan plot across the rang
6. means multiplied by e SimpliNano will default to factors 50 for double stranded DNA 40 for RNA and 33 for single stranded DNA and Oligonucleotides It also allows manual compensation for dilution aided by a dilution calculator e Nucleic acids extracted from cells are accompanied by protein and extensive purification is required to remove the protein impurity The 260 280 nm Absorbance ratio gives an indication of purity however it is only an indication and not a definitive assessment Pure DNA and RNA preparations have expected ratios of 2 1 8 and 2 2 0 respectively Deviations from this indicate the presence of impurity in the sample but care must be taken in the interpretation of results e The 260 nm reading is taken near the top of a broad peak in the Absorbance spectrum for nucleic acids whereas the 280 nm reading naturally occurs on a steep slope where small changes in wavelength will result in large changes in Absorbance Consequently small variations in wavelength accuracy have a much larger effect at 280 nm than at 260 nm It follows therefore that the 260 280 ratio is susceptible to this effect and users are warned that spectrophotometers of different designs may give slightly different ratios e n practice concentration also affects the 260 280 ratio as the individual readings approach the instrument s detection limit If a solution is too dilute the 29088793 AB 05 2014 13 280 nm reading shows a greater proportional
7. 31 10 Legal GE GE monogram and imagination at work are trademarks of General Electric Company SimpliNano is a trademark of General Electric Company or one of its subsidiaries Windows 2000 Windows XP and Windows Vista are trademarks of Microsoft Corporation in the United States and or other countries All other third party trademarks are the property of their respective owner 2013 2014 General Electric Company All rights reserved First published October 2013 All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them A copy of these terms and conditions is available on request Contact your local GE Healthcare representative for the most current information http www gelifesciences com GE Healthcare UK Limited Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Bio Sciences AB Bj rkgaton 30 751 84 Uppsala Sweden GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare Bio Sciences Corp 800 Centennial Avenue PO Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Japan Corporation Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan 29088793 AB 05 2014 32 Page left intentionally left blank Page left intentionally left blank Page left intentionally left blank GE Healthcare offices GE Healthcare Bio Sciences AB Bj rkgatan 30 751 84 Uppsala Swede
8. 3a and 3b Sample port area Figure 3a Figure 3b Step 2 Pipette the reference solution into the port A good reference will use the same solvent as the sample To ensure the correct placement of the solution in the port touch the pipette tip to the bottom of the sample reservoir and then slowly dispense the solution Step 3 The sample or reference should be pipetted into the centre of the port and be in contact with both port probes Take care not to introduce bubbles into the sample Figure 4a and 4b How to correctly load sample onto sample port Figure 4a Figure 4b 29088793 AB 05 2014 9 Figure 5 The images display the incorrect loading position of samples Figure 5 Figure 6 The position and shape of the drop shown in the image is the optimum position for the sample measurement to be made Figure 6 29088793 AB 05 2014 10 Step 4 To clean the the port simply wipe with tissue or a lint free cloth When cleaning the port it is ok to touch the probes Once the port is clean you are ready to proceed with your next sample measurement Figure 7 How to clean the sample port Step 5 Pipette sample into port exactly as performed with the reference as shown in figure 4a 4b and 6 5 1 Cleaning and general maintenance Before cleaning the case of the instrument switch off the instrument and disconnect the power cord Clean all external surfaces using a soft damp cloth A mild liquid detergent may be
9. 4 2 MJM 9 gre POSTS OI Table2 Limits of Detection for BSA Proteins can be determined in the near UV at 280 nm due to absorption by tyrosine tryptophan and phenylalanine amino acids The Abs280 varies greatly for different proteins due to their amino acid content and consequently the specific Absorption value for a particular protein must be determined e The presence of nucleic acid in the protein solution can have a significant effect due to strong nucleotide Absorbance at 280 nm e The Protein A280 application has three modes which can be selected depending on the extinction coefficient of the reference protein being used A For BSA the extinction coefficient is 6 7 AU E196 at Abs 280 for a 196 ww solution B For IgG the extinction coefficient is 13 7 AU E196 at Abs 280 for a 196 ww solution C For lysozyme the extinction coefficient is 26 4 AU E196 at Abs 280 for a 196 ww solution e Rapid measurements such as this at Abs280 are particularly useful after the isolation of proteins and peptides from mixtures using spin and HiTrap columns by centrifuge and gravity respectively Use of Background Correction e Background Correction at a wavelength well away from the protein peak is used to compensate for the effects of background Absorbance The procedure can adjust for the effects of turbidity stray particulates and high Absorbance buffer solutions e SimpliNano uses background correction at 340 nm by default pr
10. and deleted using the Options menu Select the method by pressing the relevant key pad number and then press the key Delete Method Press 1 to select Delete Method Select the method to be deleted using the left and right arrows Single wavelength Press to delete the method OR Cancel s to return to Favourites Methods folder Lock Method Methods Methods 1 Press 2 to select Lock Method gt Delete Method Lock Method Select the method to be locked using the left and right arrows one menoa Press the down arrow Select a pass code using the keypad numbers or left and right arrows Press to lock the method OR Cancel r3 to return to the Methods folder Unlock Method Press 3 to select Unlock Method Select the method to be unlocked using the left and right arrows Press the down arrow Enter the pass code using the keypad numbers or left and right arrows Press to unlock the method OR Cancel co to return to the Methods folder 29088793 AB 05 2014 22 6 UTILITIES Press 7 to enter the Utilities folder Summary Function Keypad number Description H OS 1 Set correct date and time Date and Time 2 Select preferred language and number format 3 Printer output options 4 Select screen layout themes and history on Adjust screen contrast amp brightness 6 Serial number and software version 6 1 Date and Time Enter the day using the keypad numbers or left and right arrows
11. and pmol ul If pmol ul is selected the factor changes to a selection table denoting the ratios of the 4 bases in the structure Press the down arrow 1 000 Step 5 units not pmol ul i I Enter the factor using the keypad numbers Default value is 33 range is 0 01 to 9999 OR Step 5 units pmol ul Enter the proportions of bases present using the keypad numbers and up and down arrows to move between boxes Default is 10 for each range is 0 to 9999 Step 6 Press OK to enter the Results screen and start making measurements OR co to return to the Home page Results Screen Step 7 Pipette the reference sample into the sample port Press Key This reference will be used for all subsequent samples until changed If QA is switched on the reference sample will need to be replaced and the key pressed again Step 8 Clean the sample port pipette the sample and press p This measures at the selected wavelengths and displays the results The ratio of wavelengths 1 and 2 absorbencies are calculated both corrected by the background wavelength value If selected Gives concentration based on absorbance at wavelength 1 Repeat step 8 for all samples Press Lr to return to the Home page Press ES to display the following options Options select using keypad numbers Return to Parameters screen step 1 above 2 SEE RN Q Print result via selected method Graph
12. check pipette calibration e Concentration of sample too low or too high e Particulates in sample Absorbance measurements will not be accurate in turbid samples e Possible noise or measurement stability issue Contact technical support e Check all sample paths are clear and clean dried on DNA Protein sample on the Micro volume head may cause start up calibration errors e Check original 18 V dc supply is connected and is fully engaged e Report persistent failures to technical support e You may be keeping you finger on the ON OFF button too long so that the instrument receives both ON and OFF signals and switches off after the calibration e Try adjusting the timing of your finger press at switch on e f the problem persists please contact your supplier Remember that the Absorbance displayed is being normalized to a path length of 10 mm With a 0 5 mm path length the ideal measurement range becomes equivalent when normalised to 2 A to 50 A For unresolved Absorbance issues contact technical support If the lamp can be heard buzzing at startup but there is no image on the screen then the display may be faulty Please contact your supplier 28 8 1 Frequently asked questions DO 2O DO o gt O gt O O o o What is the Pathlength of SimpliNano 0 5 mimi How accurate is SimpliNano Typically within 296 Does SimpliNano produce results for a continuous spectrum or just selected w
13. choose settings 29088793 AB 05 2014 6 e When the Auto Print option is set to OFF all data will automatically be saved to Printer the USB memory stick The operator can choose either the TSV or PVC format by using the arrows keys to select USB Stick Output PVC files can be opened NE in the PVC program TSV files can be opened in Microsoft Excel el OK TH Caca e When a USB stick is inserted and an application selected an icon will appear on the top right hand side of the status bar AZJ3UO Sample AZEU U AZO A320 Concentration ACBEOJATSDO A260 A230 Units pa mil e When the USB stick is being written to the icon will change as shown A230 Sample A260 A280 A320 Concentration A260 A280 A260 A230 Units ug ml Multiple samples are stored in the same file on the USB stick during a sample batch the icon will be displayed as shown to indicate that a file on the USB stick is j A230 000 Sampie open but not currently being written to ee eee A280 o00A A320 0 000 A Concentration A260 A280 0 00 9 A260 A230 Unis ug rn The USB stick should not be removed when this icon is displayed To safely remove the USB memory stick after a reading close the application by first pressing the A30 aoa Sample ESC key and withdraw the stick from the port WARNING DATA MAY BE LOST IF Unsafe removal of USB memory EB THE USB STICK IS REMOVED I
14. your supplier under part number 28918226 Step 1 Keep the power on Lift off the printer cover Step 2 Lock the platen horizontal position and feed the paper into the slot The drive will engage automatically and take up the paper Step 3 It may help to release the platen lock turn flat green catch clockwise and turn the green knob manually Step 4 Replace the cover 29088793 AB 05 2014 12 4 APPLICATIONS SUMMARY Function Keypad Description SimpliNano Number A e 6 1 Concentration and purity check for DNA samples A Ote 2 Concentration and purity check for RNA samples 3 Concentration and purity check for Oligonucleotide samples Oo IT 5 Concentration measurement at a single wavelength based on a factor entered or calculated from a A A single standard 5 Concentration and purity check for protein samples 6 Saved methods Instrument set up options 1 Nucleic Acid Theory Nucleic acids can be quantified at 260 nm because at this wavelength there is a clearly defined maximum peak A 50 ug ml DNA solution a 40 ug ml RNA solution and 33 ug ml solution of a typical synthetic Oligonucleotide all have an optical density of 1 0 A in a 10 mm pathlength cell These factors 50 40 and 33 respectively can be inserted into the formula 1 below although they do vary with base composition and this can be calculated more precisely if the base sequence is known 1 Concentration Abs260 Factor where
15. 1 me 9 00 5 0 s 0 gt om w aw an gt ont w Led an om 9 a oom 5 om oom oen not gt 00 oom 6 0 v 91 0 on me eo 59 2 Select the file right mouse click and select Open With from the Menu and select Microsoft Excel Data will then open directly into Excel m ewm iae Cr E dme cem a a re ETET e pe kh 9 Fi Ff A oa Ej an je 9 5 8 Lo eh O88 Oe eio Bs s wA onem m REM E jo m i ee Bind iL e jo a ee eem B enm Ame deo Di IE i imp ol E P T e y amp tems ii piit F ML mieie a Joh Ada mum s Eh Gp a Ced Lj Aei Migs am oe MP ami 1854 1 b Gm va l aka m Bi Uwe Dam ee a U see ees li ieee prea Th Feat FB ometi tate pat a i kage e Bo henge oo dearum Ar ELEI du tame e gn Jhi ane ae i B ieee L1 B cum e Li BR tem ecd ek ae P tte w wi W M Lo ALI D D DAL IALLILIL Il M M I 0 M Ana i oma DT d ada LE LTD A DLL L SD krime t m B mma Hem Li i a me P a L LI Wa LE wt pasa bas P 7 DE UI 1 L n i LJ Fire crm pm i a 29088793 AB 05 2014 8 5 SAMPLE HANDLING SimpliNano uses a simple sample port which enables users to quickly and accurately measure samples without any moving parts The sample port comprises of 2 fibre optics which are fixed 0 5 mm apart The sample should be loaded into this space Step 1 Before measurement inspect the sample port to ensure it is clean Figure
16. AB 05 2014 29 gt O PO PFO What volume of sample can use with the SimpliNano Volumes as low as 1 ul can be used A sample volume of 2 ul is recommended to minimize pipetting errors affecting results What calibration does the SimpliNano require The SimpliNano has no moving parts so calibration is not required How reproducible is the Absorbance data Specifications for precision and accuracy are below Absorbance Precision Between 0 amp 1 A 0 005 A or 196 of measurement whichever is greater 1to 24A 296 of reading above 2 A 596 of reading Absorbance Accuracy 196 at 257 nm Potassium Dichromate solution 0 7 0 8 A What are the detection limits for SimpliNano For dsDNA the range is 4 0 ng ul 2500 ng ul and for BSA 0 12 mg ml 50 mg ml What sort of reproducibility should expect with the SimpliNano Typically x 2 ng ul for dsDNA samples 100 ng ul 2500 ng ul and 2 for samples gt 100 ng ul For BSA typically 0 1 mg ml for samples 1 50 mg ml and for gt 1 mg ml tolerance of 2 What purity ratio measurements are available on the SimpliNano The SimpliNano measures 260 280 and A260 A230 ratios for Nucleic Acids and A260 A280 ratios for proteins 29088793 AB 05 2014 50 9 SPECIFICATION AND WARRAN I Y Specification Wavelength range Monochromator Wavelength calibration Spectral bandwidth Wavelength accuracy Wavelength reproducibility Light sources
17. Detector Photometric range Photometric linearity Photometric reproducibility Stray light Zero stability Noise Digital output Dimensions Weight Power input 190 1100 nm Flat Grating Automatic upon switch on 5nm 2 NM 1 nm Pulsed xenon lamp 1024 element CCD array 0 300 to 2 500 A O to 199 T 0 005 Abs or 1 of the reading whichever is the greater 546 nm 0 003 Abs 0 to 0 5 Abs 0 007 Abs 0 5 1 0 Abs lt 0 5 at 220 nm and 340 nm using NaNO 0 01 Abs hour after 20 min warm up 340 nm 0 005 pk to pk 0 002 RMS USB B port for connection to PC 260 x 390 x 130 mm 4 5 kg 18 Vdc from a 90 250 V 50 60 Hz Max 30 VA mains power pack Specifications are measured after the instrument has warmed up at a constant ambient temperature and are typical of a production unit As part of our policy of continuous development we reserve the right to alter specifications without notice Warranty e GE guarantees that the product supplied has been thoroughly tested to ensure that it meets its published specification The warranty included in the conditions of supply is valid for 24 months only if the product has been used according to the instructions supplied GE can accept no liability for loss or damage however caused arising from the faulty or incorrect use of this product e The lamp is warranted for 3 years if the product has been used according to the instructions supplied 29088793 AB 05 2014
18. Factor screen see second parameter screen to the right Enter the volume of the sample using the keypad numbers range 1 00 to 9999 Press the down arrow Enter the volume of the diluent using the keypad numbers range 1 00 to 9999 Press OK to calculate the dilution factor and return to the DNA Parameters screen OR press Cancel to cancel the selections and return to the DNA Parameters screen Step 3 Background Correction at 320 nm will be as default It may be turned off with the left or right arrows Press the down arrow Step 4 Select the units of measurement using the left and right arrows Options ug ml ng ul ug ul Press the down arrow Step 5 Enter the factor using the keypad numbers Default value is 50 the range is 0 01 to 9999 Step 6 Press OK to enter the Results screen and begin taking measurements OR Ese to return to the Home screen Results Screen Step 7 Pipette the reference sample into the sample port Press Key This reference will be used for all subsequent samples until changed If QA is switched on the reference sample will need to be replaced and the key pressed again Step 8 Clean the sample port pipette the sample into the port Press y This measures at the selected wavelengths and displays the results Repeat step 8 for all samples Press Esc to return to the Home page OR press to display the following options 29088793 AB 05 2014 15 1 000 50 0 On
19. K to enter the Results screen EB Cancel OR a s nemore Cancel Esc to return to the Protein screen Results Screen Step 8 Pipette on the reference sample and press the key This reference will be used for all subsequent samples until changed If QA is switched on the sample will need to be replaced and the key pressed again Step 9 Clean the sample port pipette the sample and press 4 This measures at the selected wavelength and displays the results Repeat step 9 for all samples Press to display the following options Options select using keyoad numbers 4 Parameters Return to Parameters screen step 1 above Q Fin r Print result via selected method Graph Toggle Graph on off The graph shows a wavescan plot across the range 250 nmto 550 nm Sample Number add a prefix to the sample number and reset the incrementing number to the desired value Sample Number Save Method Awuto Print Save Method use the alpha numeric keys to enter a name for the method and press Save Auto Print toggles Auto Print on off Exit Options by pressing r3 or wait oo 29088793 AB 05 2014 21 5 METHODS FOLDER fa m Press 6 to enter the Methods folder This folder is the storage locations for any user modified applications Methods that are saved using the Options menu and is accessible from the Home page Up to 9 Methods may be stored saved methods can be locked unlocked
20. NCORRECTLY This will be indicated by the following WP mH notification AX e HII the application before removing the USE memory stick e OK t ua nmi 29088793 AB 05 2014 7 Data transfer to USB can be carried out simultaneously with printing to a built in printer if fitted or export to a PC running PVC software if connected e Totransfer data simultaneously to PC enter Printer options screen use arrow buttons on keypad to scroll th change settings as follows e Auto Print setting ON e Printer Computer USB e USB Stick Output TSV or PVC e Totransfer data simultaneously to Printer enter Printer options screen use arrow buttons on keypad to scroll th change settings as follows e Auto Print setting ON e Printer Built in e USB Stick Output TSV or PVC 2 4 Working with data stored on a USB stick For data stored in PVC fromat please refer to the PVC user manual or help file within PVC software For data stored in TSV format insert the USB stick into a PC and use Windows Explorer to navigate to the saved file which will be located in a folder named as the instrument serial number The file can then be opened in several ways 1 Double clicking on the txt file will open the file into Windows Notepad as follows ae P tert tpm evette ww c e 279 v2 jets he iew wate it eee 034 e ww EL aee atte Pant ume I M I4 99 1 19 u
21. NG Negative absorbance readings Unexpected results Absorbance higher than expected Absorbance lower than expected Poor reproducibility Instrument start up reported failure Instrument switches off after calibration Absorbance readings stable but different than expected No image on screen 29088793 AB 05 2014 Sample measurements will be negative absorbance reading if the absorbance value of the reference is higher than the sample Negative readings can also result if reference and sample are interchanged or if the sample is very dilute and close to the absorbance of the reference Contact your supplier for advice on the minimum concentrations that can be measured e Bubbles or contamination in the sample or reference can result in considerable errors e Use of incompatible buffers UV Absorbing as a reference e Ensure background correction is switched on e Trying to measure sample volumes concentrations outside of the instrument working ranges e Incorrect sample reference e Contamination in sample e Check size and position of droplet e Ensure background correction is switched on e Possible incorrect optical alignment Contact technical support e Incorrect sample reference e Check sample and reference for contamination e Check sample and reference samples are not the same e Check size and position of droplet e Possible stray light issue Contact technical support e Insufficient sample volume
22. SimpliNano Product User Manual Codes 29061711 SimpliNano 29061712 SimpliNano with printer 29061713 SimpliNano North American version 29061714 SimpliNano with printer North American version Page finder 1 INSTALLATION 1 1 Unpacking and positioning 1 2 Safety 1 5 Declaration of conformity 2 INTRODUCTION 2 1 Your Spectrophotometer 2 2 File system 2 5 Data export 2 4 Working with data stored on a USB stick 3 SAMPLE HANDLING 5 1 Cleaning and general maintenance 3 2 Return for repair 3 3 Lamp replacement 3 4 Replacing the printer paper 4 APPLICATIONS 4 1 Nucleic Acid Theory 4 2 DNA measurement 4 5 RNA measurement 4 4 Oligo 4 5 Absorbance Concentration 4 6 Protein determination at 280 nm 4 7 Protein A280 5 METHODS FOLDER 6 UTILITIES 6 1 Date and time 6 2 Regional 6 5 Printer 6 4 Preferences 6 5 Contrast 6 6 About 7 ACCESSORIES INSTALLATION 7 1 Printer installation 7 2 After Sales Support 8 TROUBLESHOOTING 8 1 Frequently Asked Questions 9 SPECIFICATION AND WARRANTY 10 LEGAL 29088793 AB 05 2014 WO O0 Oo U0101 Ui f WNW CJ b gt be bP BS NO No Pe RN NH mn BuIDwmugDwntmpD ntmtpD CO QO Oo J DOW W MNO MNO PO o no nono n o no BORD BWW WG DMO PO no NW Ui N PO WO WN CJ N FR 1 INSTALLATION 1 Unpacking and positioning It is recommended that users read through this manual prior to use e Remove the instrument from its pac
23. an Excel spreadsheet an EMF graphics file a comma delimited csv data file a tab delimited txt data file or in native PVC format Some users may find it convenient to use PVC to print through the PC directly to a local or networked printer Datrys Computer control software is also available for SimpliNano as an optional extra The SimpliNano USB has the option to save data onto a USB stick in 2 different formats both formats save Method details all measured results and all scan data if available in the application 1 PVC format that can only be opened by Print via Computer software from GE Healthcare Once a file has been opened in PVC it can be exported into ASCII comma or tab separated text Extensible meta format or Excel format only if Excel is installed on the PC 2 Tab Separated Variable TSV allowing the data to be opened directly into Excel Please note that the USB stick needs to be placed into machine BEFORE measurements are made and encrypted sticks amp external hard drives are not compatabile with this instrument e nsert USB memory stick into the connector on the right hand side of the instrument An audible click will be heard as the stick is recognised e On main menu screen choose Utilities option by pressing 7 on the keypad e Then choose Printer option by pressing 3 on the keypad to choose Data Export Options Data output can then be made from the Printer screen Use arrow keys to scroll down and
24. and proteins 2 2 File system After switch on and automatic checking the instrument defaults to a home page entitled SimpliNano This page displays seven folders which form the topmost layer of a simple file tree which is the basis of the user interface The folder screens are reached by pressing the appropriate number on the keypad with return to the top level by means of the Esc key The folders group various facilities together as follows 1 DNA Concentration and purity check for DNA samples 2 RNA Concentration and purity check for RNA samples 3 Oligo Concentration and purity check for Oligonucleotide samples 4 Absorbance Concentration measurement at a single wavelength based on Concentration a factor entered or calculated from a single standard 5 Protein A280 Concentration and purity check for protein samples 6 Methods Saved methods 7 Utilities Instrument set up options 29088793 AB 05 2014 s 2 5 Data export An integral printer is available for the instrument this may be either supplied pre installed or as an optional accessory The installation procedure is described in section 7 1 rr d Figure 2 SimpliNano with Printer Data can be transferred to a PC via a USB cable using the supplied software PVC Print Via Computer PVC can store data either in a common directory or can be configured to save to independent directories by both file format and connection The data may be stored as
25. avelengths Within The DNA RNA Oligo and protein A280 methods a continuous spectrum is produced Do nucleic acids require purification before being measured Yes Absorbance measurements are not specific for a particular nucleic acid and will be affected by the presence of other molecules which absorb at 260 nm How should the sample port be cleaned The sample port is easy to clean Simply wipe with a lint free tissue or cloth If a difficult or sticky sample or dye was used the port can be cleaned with a dilute 296 detergent solution followed by water or isopropanol See Section 3 1 Cleaning and General Maintenance Step 4 for details Isa simple wipe with a lint free tissue or cloth enough to prevent carryover Yes A sample carryover study using 25 mg ml BSA with successive alternative additions of water and BSA was repeated 50 times The data showed no significant change in absorbance readings throughout the study for the water samples or the BSA samples indicating that there was no carryover See SimpliNano Datafile 29 1028 98 for details For more difficult sticky or strongly coloured samples the port can be cleaned with a dilute 296 detergent solution followed by water or isopropanol Are there any solvent restrictions Most common laboratory solvents including dilute acids may be used but they should be immediately wiped off the sample head after measuring The SimpliNano has been tested w
26. e 220 nm to 320 nm with cursors denoting 230 260 280 and if Background Correction selected 320 nm Sample Number Save Method Auto Print Sample Number add a prefix to the sample number and reset the incrementing number to the desired value Q Save Method use the alpha numeric keys to enter a name for the method and press Save e Auto Print toggles Auto Print on off Exit Options by pressing Esc or wait oco 4 4 Oligo The procedure is as follows Fathlenath Units step l Press 3 to select Oligo mode Dilution Factor Factor Step 2 dilution factor known Enter the dilution factor using the keypad numbers range 1 00 to 9999 Use the Faas C button to backspace and clear the last digit entered On OR Step 2 calculate Dilution Factor Press to enter the Dilution Factor screen Enter the volume of the sample using the keypad numbers range 1 00 to 9999 Parameters Press the down arrow Enter the volume of the diluent using the keyoad numbers range 1 00 to 9999 Press OK to calculate the dilution factor and return to the Parameters screen OR press co to cancel the selections and return to the Parameters screen Step 3 Select whether the background correction at 320 nm is used or not with the left and right arrows Press the down arrow 29088793 AB 05 2014 17 Step 4 Select the units of measurement using the left and right arrows Options ug ml ng ul ug ul
27. function toggles On and Off with either left right arrows from the relevant page e When background correction is on the results will be different from when it is off as Abs320 is subtracted from Abs260 as shown in the equations below Abs260 Abs320 Factor Abs260 Abs320 Abs280 Abs320 Concentration Abs ratio 260 280 Abs ratio 260 230 Abs260 Abs320 Abs230 Abs320 Abs PURE NUCLEIC ACID POLY dAdT 1 50 1 25 1 00 0 75 Wave 260 0 Abs 0 700 0 50 N Wave 280 0 Abs 0 383 0 25 4 0 00 200 0 225 0 250 0 275 0 300 0 325 0 350 0 Wavelength Typical spectral scan of a Nucleic Acid Note e The Absorbance maximum near 260 nm and Absorbance minimum near 230 nm e The flat peak near 260 nm and steep slope at 280 nm e There is very little Absorbance at 320 nm Sample Lower limit Upper Limit Reproducibility Based on 10 replicate Type of Detection of Detection measurements lt 100 ng ul Tolerance of 2 ng ul dsDNA 4 0 ng ul 2500 ng ul 100 ng 2500 ng ul Tolerance of 2 a Table 1 Limits of Detection for DNA 29088793 AB 05 2014 14 4 2 DNA measurement The procedure is as follows Step 1 Press 1 to select DNA Parameters mode Step 2 dilution factor known Enter the dilution factor using the keypad numbers range 1 00 to 9999 Use the C button to backspace and clear the last digit entered OR Step 2 calculate dilution factor Press to enter the Dilution
28. interference from background and as the divisor becomes smaller has a disproportionate effect on the final result It is advisable to ensure that the Abs260 value is greater than 0 1 A for accurate measurements e An elevated Absorbance at 230 nm can also indicate the presence of impurities 230 nm is near the Absorbance maximum of peptide bonds and may also indicate interference from common buffers such as Tris and EDTA When measuring RNA samples the 260 230 ratio should be gt 2 0 A ratio lower than this is generally indicative of contamination with guanidinium thiocyanate a reagent commonly used in RNA purification which absorbs over the 250 260 nm range A wavelength scan of the nucleic acid is particularly useful for RNA samples e SimpliNano displays the individual Absorbance values and the 260 280 and 260 230 ratios on the left side of the screen and attention should be given to these as well as the final result on the right hand side Use of Background Correction e Background Correction at a wavelength well away from the nucleic acid or protein peaks is often used to compensate for the effects of background absorbance The procedure can adjust for the effects of turbidity stray particulates and high Absorbance buffer solutions e SimpliNano uses background correction at 320 nm by default for nucleic acid measurements It is particularly recommended since very small samples are susceptible to stray particulates The background
29. ith methanol isopropanol 2 propanol and acetone However these solvents may affect the accuracy of the measurement due to their volatility amp are therefore not recommended Why does the reference blank require 2 samples To set the reference the software will ask for 2 separate samples to ensure the quality of the reference baseline instrument What kind of light source does the SimpliNano use A pulsed Xenon flash lamp How often will the lamp require changing The lamp should last for several years of use amp comes with a 3 year warranty If it does need changing this should be done by a service engineer from your supplier Is the flash lamp on continuously or only when performing a measurement The lamp is only on when taking measurements Does SimpliNano require a computer to operate No SimpliNano operates as a stand alone instrument and data can be transferred via USB stick If operation via a computer is critical it can be connected to a PC and operated using either PVC or Datrys software Can get data exported in Excel format Using the PVC software application data may be stored either manually or automatically in a variety of data formats which include as an Excel spreadsheet Excel must be installed on the PC Extensible Meta Format an EMF graphics file a comma delimited csv data file a tab delimited txt file Rich Text Format RTF or in native PVC format 29088793
30. kaging and inspect it for signs of damage If any are discovered inform your supplier immediately e The instrument must be placed on a stable level surface that can take its weight 4 5 kg and positioned such that air can circulate freely around the casing e Ensure your proposed installation site conforms to the environmental conditions for safe operation e The instrument is designed for indoor use only temperature range 5 C to 35 C and should be kept away from strong draughts e f you use the instrument in a room subjected to extremes of temperature change during the day it may be necessary to recalibrate by switching off and then on again once thermal equilibrium has been established 2 3 hours e Atemperature change of no more than 4 C hour and a maximum relative humidity of 80 at 31 C decreasing linearly to 50 at 40 C are required e fthe instrument has just been unpacked or has been stored in a cold environment it should be allowed to come to thermal equilibrium for 2 3 hours in the laboratory before switching on This will prevent calibration failure as a result of internal condensation e The instrument must be connected to the power supply with the power adaptor supplied The adaptor can be used on 90 240 V 50 60 Hz supplies It will become warm once plugged into the power supply and should not be covered up e Switch on the instrument via the keypad after it has been plugged in The instrument will pe
31. mple and press 4 This measures at the selected wavelength and displays the results Repeat step 7 for all samples Step 8 if Standard selected Insert standard and press Change the concentration value of the standard If necessary and press ok ES to measure the standard Insert sample and press This measures at the selected wavelength and displays the results Repeat for all samples Press E to display the following options Press Cancel Esc to cancel selections and return to the Home page Options select using key pad numbers Return to Parameters screen step 1 above Print result via selected method Run Standard Sample number add a prefix to the sample number and reset the incrementing number to the desired value Save method use the alpha numeric keys to enter a name for the method and press Save Auto print toggles auto print on off Exit options by pressing esc or wait 29088793 AB 05 2014 19 l Absorbance Concentration ES 5 0 0 Factor e 1042 Units 130 nm Standard Parameters e Prit Q Run Standard Concentration 90 0 save Method Units O Ato P rint 4 6 Protein determination at 280 nm Sample Lower limit Upper Limit Reproducibility Based on 10 replicate Type of Detection of Detection measurements 1 mg ml Tolerance of 0 06 mg ml BSA 0 12 mg ml 50 mg ml PN ml Tol
32. n GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Japan Corporation Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan For contact information for your local office please visit www gelifesciences com contact GE Healthcare Limited Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK http www gelifesciences com imagination at work 29088793 AB 05 2014
33. otein measurements It is particularly recommended since very small samples are susceptible to stray particulates The Background function toggles On and Off with either left right arrows from the relevant page e When background correction is on the results will be different from when it is off as Abs340 is subtracted from Abs280 prior to reporting 4 7 Protein A280 Step 1 Press 5 to select Protein A280 Step 2 Select the Mode Options are BSA IgG Lysozyme Press the down arrow Step 3 dilution factor known ok G Cancel Enter the dilution factor using the keypad numbers range 1 000 to 9999 Use the C button to backspace and clear the last digit entered 29088793 AB 05 2014 20 OR Step 4 calculate dilution factor Press to enter the dilution factor screen shown to the right Enter the volume of the sample using the keypad numbers range 0 001 to 9999 Press the down arrow Enter the volume of the diluent using the keyoad numbers range 0 001 to 9999 Press OK to calculate the dilution factor and return to the Parameters screen OR press co to cancel the selections and return to the Parameters screen Step 5 Select whether Background Correction is to be used or not with the left and right arrows i Background Press the down arrow On Step 6 Pathlength Select the Units of measurement using the left and right arrows Options mg ml ug ml ng ul and ug ul Dilution Factor Step 7 Press O
34. p LI eee emg o gt to 9 e LASI ws el o t 1 e tu 9 oo wr oe ei IM 70 ioa wt n w ne LIE 41 ee 221 eee ee oe LE E ace oe o LE es tt su 4 t LE SA oe t e e os LE A EL et ul ve ad 1o J 14 7r B e wn v ww e om 4 9 LM b own o Gu gt en 4 wet c4 61 o gt oe o ooi oom 2 oog o O6 0 Goo 6 ome oot o oj 5 00 oo E 20 1 5 e cue 5 qu 0 9 D eU 5 000 6 5 ae tuno 0 om 6 ow 6 cM 0e 0 6 S mw xd 2 oni gt ow 9 m 6 o gt e e e one o d 0 we IM oa n oe c E o 6 o o So gt U ow o0 m Umm 3 oe o que UL ej 6 66 0 GU o o U wo o dw a gt om wa E e Amp n oe e 5 0M 061 e o 5 aw E cnn me oot wo wn on gt n gt wt M qe 6 et 0 e b omn A l EN ae gt oso wi mtd 1 wi me 9 wi mi w E e wx LI gt ons 6 00 oot 1 bow e ool o en we n o4 wj mi omy gt oh b 0M o4 oue 1 wo LM 6 oc 6j 6 X ww a oon Ld gt we mw m4 wo Iu ont gt wu Dll ILI ww a o O1 gt qu ene G1 o9 Daw Cane r n1 0 c o9 eo e Gh e Ga m uu i ow a Lo 9 o0 on i LI e wi w oor am wo 691 001 5 0 5 0M Ln _ P o LIU ao t gt m DI o gt Ow de IL on wo i 56 qu gt aM op ort 1 d e we LUI D 9 w o ou w e o0 e qu we o an e os e wc 0 oot o Gc gt w mec 5 om 9 E al LL td 9 wi 001 Xx 6 Oi MELZ eo oD oa 90 06 e om e w e 5 Li A 4 0
35. requirements of the following Directives 2006 95 EC Low voltage directive LVD 2004 108 EC Electromagnetic Compatibility EMC directive 2012 19 EU Waste Electrical and Electronic Equipment directive recast WEEE Recast 2011 65 EU Restriction on the use of certain hazardous substances ROHS directive 2006 42 EC Machinery directive Standards to which conformity is declared are as follows EN61010 1 2010 Safety requirements for electrical equipment for measurement control and laboratory use General requirements EN61326 1 2006 Electrical equipment for measurement control and laboratory use EMC Requirements EN50581 2012 Technical documentation for the assessment of electrical and electronic products with respect to the restriction of hazardous substances EN ISO 12100 2010 Safety of machinery General principles for design risk assessment and risk reduction 29088793 AB 05 2014 4 l INTRODUCTION 2 1 Your Spectrophotometer SimpliNano is a simple to use UV Visible instrument with twin CCD array detectors 1024 pixels It has no moving parts which Is the basis of the rapid scanning operating system Display On Off key USB A connector USB B connector to PC Power input Sample Port Figure 1 SimpliNano Spectrophotometer The SimpliNano is fitted with a micro volume sampling port with a fixed 0 5 mm pathlength for measuring low volumes of highly absorbing life science samples such as DNA RNA
36. rform a series of self diagnostic checks e Contact your supplier if you experience any difficulties with this instrument 1 2 Safety Spectrophotometer Health amp Safety Document including General Operating Instructions are available as a booklet provided with each instrument The booklet is translated into the European languages and is available on the User Manual CD supplied with each instrument The instructions cover basic operation troubleshooting and how to use the instrument in a safe manner A CAUTION The instruments contain a UV source which generates a light beam that traverses the sample chamber and is accessible in the lamp chamber Under normal use the lamp beam is confined within the instrument The unit should not be operated with the lamp housing lid removed as prolonged exposure to the beam may cause permanent eye damage A WARNING High voltages exist inside the SimpliNano instruments Repair and maintenance should only be carried out by individuals trained specifically to work on these instruments 29088793 AB 05 2014 3 If the instrument is used in a manner not specified or in environmental conditions not appropriate for safe operation the protection provided may be impaired and instrument warranty withdrawn There are no user serviceable parts inside this instrument 1 3 Declaration of conformity GE certifies that SimpliNano UV Visible Spectrophotometers part numbers 29061711 29061712 conform to the
37. screen without storing the settings 6 4 Preferences Preterences Sets user Preferences EE ZEN Quality Assurance On Define the screen layout of folders Options are either a grid format default or a list Press the down arrow Select whether to use previously entered parameters when the instrument is switched on or to use defaults Press the down arrow e o Cane Select whether to use a standby mode after defined periods Options are 1 hour 2 hours at night or off Select whether you want Quality Assurance mode on off QA mode prompts you to run a 2nd Reference and checks that both references agree within a specified tolerance Press OK p to store the settings and return to the Utilities screen OR Press Cancel Esc to return to the Utilities screen without storing the settings 6 5 Contrast Ambient temperature can affect the display This function can optimise the display for local conditions The procedure is as follows Adjust the contrast using the left and right arrows to scroll though the levels Press the down arrow briefly to move down Adjust the brightness similarly Press the down arrow Press OK to store the settings and return to the Utilities screen 6 6 About Displays the instrument serial number and software version Press OK to close the window and return to the Utilities screen Serial Number 54321 Version 4290 v1 6 e OK 29088793 AB 05 2014 24
38. used to remove stubborn marks The sample port can be cleaned as follows e tis recommended that before starting the sample port is wiped with a lint free tissue dampened with distilled water e For routine use wiping the sample port with a dry lint free tissue between samples is sufficient to remove sample e Periodic cleaning of the sample port is recommended after around 100 samples or if using sticky protein samples In this case a quick wash with a laboratory cleaning detergent is sufficient to remove build up from proteins and DNA Remove detergent by pipetting on distilled water maximum volume 25 ul and removing with a lint free tissue e Use of spray or squeezy bottles should be avoided as these flood the sample area 3 2 Return for repair The responsibility for decontamination of the instrument lies with the customer The case may be cleaned with mild detergent or an alcohol such as ethanol or Isopropanol Examination or repair of returned instruments cannot be undertaken unless they are accompanied by a decontamination certificate signed by a responsible person This could be of the following form 29088793 AB 05 2014 11 3 3 Lamp replacement The Xenon lamp should not need replacement until used for several years In the unlikely event that it fails this should be undertaken by a service engineer from your supplier 3 4 Replacing the printer paper Paper for the printer 20 rolls is available from
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