Qubit® 2.0 Fluorometer - Thermo Fisher Scientific
Contents
1. 0 100 200 300 400 500 Concentration ng mL e A Standards Incorrect message indicates an error Checking the calibration values may help you to determine the source of your error See Checking Standards next page w 19 Mar AHO ILIO PM vZ dsDNA HS O 100 200 300 400 So Goncenbration ng mL Read New i Standards Home Standards Using the Last You can choose to apply the previous calibration to your sample Calibration readings by pressing No when the Qubit 2 0 Fluorometer prompts you to choose between reading new standards and using the previous calibration You are then directed to insert the assay tube containing your sample Continued on next page 17 Calibrating the Qubit 2 0 Fluorometer Continued Checking Standards The Check Standards Screen displays the raw fluorescence values for Standard 1 Standard 2 Standard 3 if applicable and the last measurement taken for a given assay These values can assist you in making a judgment regarding the performance of the Qubit 2 0 Fluorometer and whether or not the samples you used were within range For the Qubit dsDNA BR Qubit dsDNA HS Qubit ssDNA Qubit RNA and Qubit RNA BR assays the reading given by Standard 2 should be much higher than that of Standard 1 and the reading of your sample should fall somewhere between that of the two Standards For the Qubit protein assay the reading given by Standard 3 should be mu2. 2 0 Fluorometer is shipped with the components listed below Component Quantity Oubit 2 0 Fluorometer 1 Universal Power Cord with 4 adaptor plugs 1 for U S E U U K Australia Qubit 2 0 USB drive 1 includes the instruction manual Oubit 2 0 Fluorometer Quick Reference Card 1 QRC Certificate of Conformity COC 1 The complete instruction manual is also available on our website at www invitrogen com qubit See pages 5 8 for the description and specifications of the Qubit 2 0 Fluorometer Examine the instrument carefully for damage incurred during transit Ensure that all parts of the instrument including accessories listed above are included with the product Damage claims must be filed with the carrier the warranty does not cover in transit damage See page 9 to install the instrument Visit probes invitrogen com qubit to register your instrument You will be asked to supply the serial number your name and your contact details Registering your instrument ensures that you will receive notifications of software upgrades and information on new assays for use with the Qubit 2 0 Fluorometer For research use only Not intended for any animal or human therapeutic or diagnostic use Safety Information Safety Precautions Review and follow the safety instructions below Do not install the instrument in heavy humidity such as ina greenhouse or an incubator to avoid a danger of electric shock If wa
3. hides the Fluorescence vs Concentration graph and the screen only displays the latest measurement Note that the Hide Graph function is not available on the Standards Screen G is Mac 200 PM an dsDNA HS Assay Tuoe Conc E o Rese Next Caloulate Sample Stock Conc Home Standards Pressing Show Graph brings the graph back onto the screen Eee J 100 700 3200 400 500 Concentration ngimk Assay Tube Conc ME on Read Next Calnulate Sample Stock Conc Home Standards Data Pressing Clear Graph removes all sample data from the Fluorescence vs Concentration graph but keeps the standards data Qubit 2 0 Fluorometer automatically stores numeric data from all reads but does not store graphic data or calculations To save all data and calculations to the Qubit 2 0 USB Drive see Saving the Data to the Qubit 2 0 USB Drive on page 27 21 Calculating the Original Sample Concentration Introduction Dilution Calculator The Dilution Calculator feature of the Qubit 2 0 Fluorometer calculates the concentration of your original sample based on the volume of sample you have added to the assay tube 1 To calculate the concentration of your original sample press Calculate Stock Conc Assay Tube Conc a oo Head Met Catmdate Tampi Stock Conc Home Standards Data The Dilution Calculator Screen containing the volume roller wheel is displayed _ 19 Mae 2010 12 30 4 E HAD
4. power down the Qubit 2 0 Fluorometer unplug the instrument Qubit 2 0 Fluorometer Loading Continued on next page Getting Started Continued Sleep Mode Home Screen The Qubit 2 0 Fluorometer has an auto standby i e sleep mode that is triggered after 30 minutes of inactivity You can also trigger the stand by mode by touching the Power button E located on the top left of the touch screen To recover from the standby mode touch anywhere on the touch screen Recovery from standby mode returns you to the same screen present when sleep mode commenced After 7 5 hours of inactivity touching the screen returns you to the Home Screen see below When the instrument is turned on the Home Screen is displayed From the Home Screen you can proceed immediately to the quantitation assays DNA RNA or protein choose to calibrate the instrument with the appropriate standards access the saved data set up the date and time or adjust the screen brightness O 19ma zomw nain wo Choose Your Assay DNA RNA Protein Continued on next page 10 Getting Started Continued Date and Time Setup Adjusting Screen Brightness The date and time is already preset when you receive the instrument To reset the date and time follow these steps 1 Press Date Time The Date Time Properties Screen contains 6 roller wheels that respond to movement of a finger across the screen as if they were wheel
5. 2008 Continued on next page 39 Purchaser Notification Continued Instrument Warranty Invitrogen a part of Life Technologies Corporation warrants to the original purchaser Purchaser that the Instrument Instrument will be free from defects in materials and workmanship for a period of one 1 year from the date of delivery Invitrogen agrees as its sole responsibility under this limited warranty and upon prompt notice of a defect to repair replace or credit the purchase price at its discretion any Instrument discovered to be defective within the warranty period Invitrogen will credit the account within 30 days of purchase if the instrument is in original condition After 30 days Invitrogen will only replace the instrument for up to a year and no credit will be issued Invitrogen will not accept any returned instrument that was used in HIV or other infectious disease labs This warranty does not include repair replacement or refund necessitated by accident abuse neglect misuse unauthorized repair or modification of the Instrument The warranty will be voided if the instrument is disassembled or a customer attempted to repair the instrument In the event that Invitrogen determines that the Instrument is in need of repair and not replacement this Standard Warranty includes replacement parts and labor for the Instrument This Standard Warranty does not include shipment of the Instrument to and from service lo
6. 3211 500 assays 033212 36 Regulatory Requirements w oN N A Q Ix The Qubit 2 0 Fluorometer has been tested and complies with the following regulatory requirements U S and Canadian Safety Standards UL 61010 1 2004 for U S CAN CSA C22 2 No 61010 1 2004 for Canada U S and Canadian EMC Requirements FCC CFR 47 Part 15 Sections 15 107 and 15 109 Class B Industry Canada Interference Causing Equipment Standard ICES 003 Issue 4 Sections 5 3 and 5 5 Digital apparatus European Safety and EMC Standards 2006 95 EC Low Voltage directive EN 61010 1 2001 EN 6101 2 081 2001 A1 2003 2004 108 EC EMC Directive EN 61326 1 2006 2002 95 EC RoHS directive 2002 96 EC WEEE directive Do not dispose in the regular household garbage Dispose the battery into the special waste for batteries Australian Directives The Australian Communication Authority ACA requirements for ISM Equipment AS NZS 2064 equivalent to EN 55011 or CISPR 11 PSE Mark for Japan Protection Class II double insulation CCC Mark for China 37 Technical Support Web Resources Visit the Invitrogen website at www invitrogen com for e Technical resources including manuals vector maps and sequences application notes MSDSs FAQs formulations citations handbooks etc e Complete technical support contact information e Access to the Invitrogen Online Catalog e Additional product information and special offers Contact Us For more i
7. 5 minutes for the Qubit protein assay e Qubit assay tubes or other appropriate 0 5 mL assay tubes e Optional Qubit 2 0 USB drive for data transfer supplied with the instrument or available separately 1 Choose Sample to go to the Sample Screen G na Mar 2000 12 30 PM dsDNA HS hictcece ha O 100 200 200 400 B00 Cenventration ing mL Insert Assay Tube Read Home Standards Continued on next page 19 Reading Samples Continued Reading 2 Insert a sample into the Sample Chamber and press Read The Samples measurement takes approximately 3 seconds continued SMa MAMA WO dsDha HS Raad Next Caloutate Sampie Initial Cone Data Upon the completion of the measurement the result is displayed on the screen The number displayed is the concentration of the nucleic acid or protein in the assay tube To calculate the concentration of your original sample see Calculating the Original Sample Concentration page 22 19 Mar 2010 12 39 P dsDNA HS _ 0 100 700 200 400 500 Concentration ng imk Assay Tube Conc BE on Read Next Calnudate Sample Stock Conc Home Standards 3 To read the next sample remove the sample from the Sample Chamber insert the next sample and press Read Next Sample 4 Repeat sample readings until all samples have been read Continued on next page 20 Reading Samples Continued Notes on Using e Graphs Pressing Hide Graph
8. Data Screen The selected line is highlighted Note By default the entire CSV file will be saved to the USB drive if no lines are selected Save your data on the USB drive by pressing the USB icon The numeric data is automatically saved as a CSV file that can be opened with any spreadsheet program Name Date 2530 15 pm 16 Aus 0 2 30 15 pm 15 Aug 2010 JASON 3 16 Aug 2010 fi JASON 3 16 Aug 2010 fi Save to USB drive complete Saving to USB drive ote fi Rename ee EH Rename 7 Home Stendands Sample Home Standards Sample To delete all data from the CSV file and start with a blank file press the Clear Data button Transfer the Qubit 2 0 USB drive to the USB port on your PC You may open the CSV file using a spreadsheet program 28 Cleaning and Maintenance Cleaning and Maintenance The Qubit 2 0 Fluorometer does not need regular maintenance To troubleshoot problems with the instrument contact Technical Support page 38 Do not perform any repairs or service on the Qubit 2 0 Fluorometer to avoid damaging the instrument e Do not expose the Qubit 2 0 Fluorometer to direct sunlight e Clean the surface of the Qubit 2 0 Fluorometer with a damp cloth e Toclean the touch screen turn off the Qubit 2 0 Fluorometer disconnect the power cable and clean the touch screen with a soft cloth lightly moistened with LCD cleansing detergent Cleaning the screen with excessive force can dama
9. NA HS Volume of Sample Used 11 pl 10 nitzal Conc a Head fect Saniple dive Home Shancdands 2 Using the volume roller wheel select the volume of your original sample that you have added to the assay tube When you stop scrolling the Qubit 2 0 Fluorometer calculates the original sample concentration based on the measured assay concentration Continued on next page 22 Calculating the Original Sample Concentration Continued Dilution 3 To change the units in which the original sample Calculator concentration is displayed press ng mL A pop up window continued showing the current unit selection as indicated by an adjacent red dash opens C 19 Mar 2010 12 30PH sv dsDNA HS Volume of Sample Used ng L Home Standards 4 Select the unit for your original sample concentration by touching the desired unit in the unit selection pop up window To close the unit selection pop up window touch anywhere on the screen outside the pop up The Qubit 2 0 Fluorometer automatically converts the units to your selection when the unit selection pop up window is closed Note The unit button next to your sample concentration reflects the change in the units e g if you change the unit to pg uL the button displays pg uL 5 To save the data from your calculation see Saving the Calculation to the Qubit 2 0 Fluorometer next page 6 To exit the Dilution Calculator Screen press any navigator b
10. The Qubit protein assay requires 15 minutes of incubation for a stable signal For greatest accuracy of the protein assay the incubation time of the samples should be within 10 minutes of the incubation time of the standards Continued on next page 34 Critical Qubit Assay Considerations Continued Photobleaching The Qubit reagents exhibit high photostability in the Qubit 2 0 of Qubit Reagents Calibrating the Qubit 2 0 Fluorometer Fluorometer showing lt 0 3 drop in fluorescence after 9 readings and lt 2 5 drop in fluorescence after 40 readings It is important to remember however that if the assay tube remains in the Qubit 2 0 Fluorometer for multiple readings a temporary reduction in fluorescence will be observed as the solution increases in temperature see Figure 2 in Assay Temperature previous page Note that the temperature inside the Qubit 2 0 Fluorometer may be as much as 3 C above room temperature after 1 hour For this reason if you want to perform multiple readings of a single tube remove the tube from the instrument and let it equilibrate to room temperature for 30 seconds before taking another reading For each assay you have the choice to run a new calibration or to use the values from the previous calibration As you first use the instrument you should perform a new calibration each time As you become familiar with the assays the instrument your pipetting accuracy and significan
11. after mixing the sample or standard with the working solution If you are performing multiple readings of a single tube remove the tube from the instrument and let it equilibrate to room temperature for 30 seconds before taking another reading Visit www invitrogen com qubit for additional application notes technical notes citations firmware updates and a list of validated Qubit assays that have been tested using the Oubit 2 0 Fluorometer Only thin wall clear 0 5 mL PCR tubes are appropriate for use in the Qubit 2 0 Fluorometer Acceptable tubes include Qubit assay tubes Cat no Q32856 500 tubes or Axygen PCR 05 C tubes VWR part number 10011 830 The minimum assay volume must be 200 uL for an accurate read 13 Calibrating the Qubit 2 0 Fluorometer Introduction Note Materials Needed Running New Standards for Calibration For each assay you have the choice to run new standards for calibrating the Qubit 2 0 Fluorometer or to use the values from the previous calibration for more information see Calibrating the Qubit 2 0 Fluorometer in the Appendix page 35 This section provides instructions for running new calibration standards and for using the last calibration Note When the fluorometer is used for the first time there will not be an option to use the last calibration Calibration of the Qubit 2 0 Fluorometer requires the preparation of the appropriate standard solutions For instructio
12. alibrating the Qubit 2 0 Fluorometer Continued Running New 4 Insert Standard 1 into the Sample Chamber and press Read Standards for Ensure that you are using the Standard 1 appropriate for the Calibration assay you are performing The reading takes approximately continued 3 seconds Reading standard 1 Insert Standard 1 Read Hore Sania CJ 19 Mw 2010 12 30 PH dsDNA HS Standard 1 complete Insert Standard 1 Read Hore Sampie Data 5 Insert Standard 2 and press Read Ensure that you are using the Standard 2 appropriate for the assay you are performing 6 If you are performing the Qubit protein assay which requires a three point calibration insert Standard 3 when prompted and then press Read The calibration is complete after Standard 2 is read or after Standard 3 is read for the Qubit protein assay The new standards graph with data points for standards connected by a line appears on the screen 16 0 100 200 300 400 500 Read New Standards No 10 1 45PM Continued on next page Calibrating the Qubit 2 0 Fluorometer Continued e Inthe Fluorescence vs Concentration graph open circles represent correct standards black circles represent samples that fall within the assay s range and red circles represent samples or standards that are not within the assay s range The most recent sample is signified by a large black circle Note Fluorescence
13. cation or travel cost of service engineer the costs of which shall be borne by the Purchaser This Warranty and the remedies set forth herein are exclusive and in lieu of all other express or implied warranties including implied watranties of merchantability fitness for a particular purpose and non infringement and no other warranties shall be binding upon Invitrogen In no event shall Invitrogen be liable for any special incidental or consequential damages resulting from the use or malfunction of this Instrument or the system with which it is used even if such damages could be anticipated by Life Technologies Corporation To obtain service during the warranty period contact Invitrogen Technical Support for further instruction OUT OF WARRANTY SERVICE Contact Invitrogen Technical Support We will be happy to assist you by phone at no charge Repair service if needed will be billed depending on the parts replaced and labor hours needed to repair your instrument You will be billed for shipment of the instrument to the recommended service facility 2010 Life Technologies Corporation All rights reserved The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners 40 amp invitrogen by Life technologies Corporate Headquarters 5791 Van Allen Way Carlsbad CA 92008 T 1 760 603 7200 F 1 760 602 6500 E tech_support invitrogen com For country specific c
14. ch higher than that of Standard 2 and the reading of your sample should be no more than 50 greater than Standard 3 To access the Check Standards Screen press Check Stds on the Standards Screen The raw fluorescence values for the standards and the last sample will be displayed on the screen C te hec mw OPM dO dsDNA HS Standard 1 Beery las ae Le Siani Creaine 19 Mar 2010 A5 P Remi New Standards Yes Home Stendands Sample Data 18 Reading Samples Introduction Note Materials Needed Reading Samples After you have completed the calibration of the Qubit 2 0 Fluorometer by running the appropriate standards for the assay you want to use or have accepted the values from the previous calibration you are ready to read your samples This section provides instructions for using the Qubit 2 0 Fluorometer to quantitate your DNA RNA or protein samples with the appropriate Qubit assay It is important to prepare the sample correctly before a reading is taken For instructions on the preparation of your samples see the instructions that accompany the assay you are using e Qubit assay kit you want to use to quantitate your samples see page 33 for available Qubit assay kits and ordering information e Your DNA RNA or protein samples Note Incubate the samples for the appropriate amount of time after mixing them with the working solution 2 minutes for the Qubit DNA and RNA assays 1
15. derations Continued Assay Temperature Incubation Time The Qubit assays were designed to be performed at room temperature 22 28 C and temperature fluctuations can influence the accuracy of the assay To minimize temperature fluctuations store all kit reagents at room temperature and insert all assay tubes into the Qubit 2 0 Fluorometer only for as much time as it takes for the instrument to measure the fluorescence because the Qubit 2 0 Fluorometer can raise the temperature of the assay solution significantly even over a period of a few minutes Do not hold the assay tubes in your hand before a measurement because holding the tubes warms the solution and results in a low reading 160 o Ma 7 140 a o 2a 120 s5 3 100 Im oOo go se ca G a SA 403 pa E 20 o 0R i i i l 10 15 20 25 30 35 Assay temperature C Figure 2 Effect of temperature on the Qubit dsDNA BR assay Qubit dsDNA HS Qubit ssDNA Qubit RNA Qubit RNA BR and Qubit protein assays show similar sensitivities over the same range To allow the Qubit assay to reach maximum fluorescence incubate the tubes for the DNA and RNA assays for 2 minutes after mixing the sample or standard with the working solution After this incubation period the fluorescence signal is stable for 3 hours at room temperature for all nucleic acid assays except the Qubit ssDNA assay which is stable for up to 30 minutes
16. ge the touch screen Wipe the screen dry immediately 29 Troubleshooting Handling Samples The calibration standards included in the Qubit RNA and Qubit RNA BR Assay Kits are high quality rRNA standards The integrity and concentration of these standards is critical to the optimal performance of the Qubit RNA assay We highly recommend treating the rRNA standards as you would any other precious RNA Use appropriate RNAse free handling techniques including RNAse free gloves pipette tips and tubes Keep the tube lids closed whenever possible do not touch the pipet to the inside wall of the tube when withdrawing a sample and return the rRNA standard to the refrigerator as soon as possible after use Ensure that the assay tubes are at room temperature at the time the reading is taken Do not hold assay tubes in your hand and do not leave assay tubes in the Qubit 2 0 Fluorometer for longer than it takes to read the fluorescence See Assay Temperature on page 34 Be careful not to spill sample into the sample chamber Promptly wipe any spills The Qubit assays are very sensitive and even small amounts of material from a previous sample may result in errors Use a clean 0 5 mL PCR tube for each reading The tube must be clean and dry on the outside when taking readings Moisture and condensation on the tube surface can lead to reading errors Minute bubbles in samples will cause errors in readings Be sure not to int
17. hat the instrument recognizes the USB drive a red dot indicates that the USB drive is not inserted into the USB port or that the instrument does not recognize the USB drive After the successful update of the firmware the following screen is displayed before the instrument returns to the Home Screen You can now proceed to setting your standards and quantitating your samples To update L Download new firmware from our website Firmware update complete Update 12 Guidelines for Using the Qubit 2 0 Fluorometer Assay Tubes for the Qubit 2 0 Fluorometer To obtain the best results follow the recommendations below For more information see Critical Qubit Assay Considerations in the Appendix pages 33 35 Do not operate the instrument in direct sunlight Wear gloves during sample handling Use the Qubit 2 0 Fluorometer at room temperature only 22 28 C Store all kit reagents at room temperature and insert all assay tubes into the Qubit 2 0 Fluorometer only for as much time as it takes for the instrument to measure the fluorescence Do not hold the assay tubes in your hand before performing a measurement Make sure that you have calibrated the Qubit 2 0 Fluorometer using the appropriate standards Incubate the tubes for the Qubit DNA and RNA assays for 2 minutes after mixing the sample or standard with the working solution Incubate the tubes for the Qubit protein assays for 15 minutes
18. invitrogen by technologies Qubit 2 0 Fluorometer Catalog no Q32866 Rev Date 4 October 2010 Manual part no MP32866 MANO0003231 Contents PROCU CEC OMLC IES erri hanes aR EA 3 Salet MMOLMATION sists atiie enactment eounieiat ela e recesses ane aaenns ee ee epaas 4 HUTFOCUICUION ioeina aaeei 5 Description of Qubit 2 0 FIUOrOMeter ssusrnnsari aiir a 5 Produch CCIM CATIONS rca A tens astuitestieses 8 Method Sasiia aaaea Ea 9 Cetine Started eraa E NA S 9 Guidelines for Using the Qubit 2 0 Fluorometer ssesseseeeeeerreeerererrsrrrrerrreereeees 13 Calibrating the Qubit 2 0 FuUorometet orrori ne a E 14 Readinie Sample Saser NAAA 19 Calculating the Original Sample Concentration ssssesesesessesesesesesrsrreesesesesereseses 22 Cleaning and WAAC CS coset anes toee tt es Saitou E E ETE 29 Troubles HOON eaa ita c cca ues caatacues ato deusast cate wseil sa waeteaton ea ovbasussieu vinmeiatentans 30 yAN 6 01 19 6 gt scini ener MEE eRe ren na ern a enter eer ner eer aa veered 33 Critical Qubit Assay Considerations ssssscssssssesescssssesssesssssessssevessseeeasennes 33 Accessory Prod UCIS sxt tst eanantee acre alae S A 36 Reculatory Requirements srir a A OA 37 Technical SU OP ORG wsimsiaa esti ses scauss exes sinn AN N 38 Purchaser NOUMCAU ON csieats Ack nn e ean 39 Product Contents Qubit 2 0 Fluorometer Upon Receiving the Instrument Registering Your Instrument Intended Use The Qubit
19. n Further the very latest illumination and detection technologies used in the Qubit 2 0 Fluorometer for attaining the highest sensitivity allow you to use as little as 1 uL of sample and still achieve high levels of accuracy even with very dilute samples Important features of the Qubit 2 0 Fluorometer are Provides a user friendly benchtop design for simple fast and highly accurate quantitation of DNA RNA and protein in less than 5 seconds per sample with sample incubation times of 2 minutes for DNA and RNA and 15 minutes for proteins Uses the Qubit assays that contain advanced dyes that only fluoresce when bound to DNA RNA or protein This specificity allows you to get very accurate results because Qubit technology only reports the concentration of the molecule of interest not contaminants Uses disposable assay tubes that eliminate washing steps and cross contamination between samples Presents comprehensive data with graphic reports and a CSV comma separated value file for sample comparisons Continued on next page Description of Qubit 2 0 Fluorometer Continued Instrument Exterior Components The image below shows the location of the various parts of the Oubit 2 0 Fluorometer Power Inlet connects the Qubit 2 0 Fluorometer to an electrical outlet using the supplied power cord and the appropriate plug based on the electrical outlet configuration in your country Touchscreen located in the f
20. nformation or technical assistance call write fax or email Additional international offices are listed on our website www invitrogen com Corporate Headquarters Japanese Headquarters European Headquarters 5791 Van Allen Way LOOP X Bldg 6F Inchinnan Business Park Carlsbad CA 92008 USA 3 9 15 Kaigan 3 Fountain Drive Tel 1 760 603 7200 Minato ku Tokyo 108 Paisley PA4 9RF UK Tel Toll Free 1 800 955 6288 0022 Tel 44 0 141 814 6100 Fax 1 760 602 6500 Tel 81 3 5730 6509 Tech Fax 44 0 141 814 E mail Fax 81 3 5730 6519 6117 tech_support invitrogen com E mail E mail jpinfo invitrogen com eurotech invitrogen com 38 Purchaser Notification Limited Use Label License No 358 Research Use Only The purchase of this product conveys to the purchaser the limited non transferable right to use the purchased amount of the product only to perform internal research for the sole benefit of the purchaser No right to resell this product or any of its components is conveyed expressly by implication or by estoppel This product is for internal research purposes only and is not for use in commercial services of any kind including without limitation reporting the results of purchaser s activities for a fee or other form of consideration For information on obtaining additional rights please contact outlicensing lifetech com or Out Licensing Life Technologies Corporation 5791 Van Allen Way Carlsbad California 9
21. ns on the preparation of these standards see the instructions that accompany the assay you are using e Appropriate standards for the assay you are performing Note Incubate the standards for the appropriate amount of time after mixing them with the working solution 2 minutes for the Qubit DNA and RNA assays 15 minutes for the Qubit protein assay e Qubit assay tubes or other appropriate 0 5 mL assay tubes e Optional Qubit 2 0 USB drive for data transfer supplied with the instrument or available separately 1 On the Home Screen choose the type of assay for which you want to run the standards Standards Screen is automatically displayed we 2wvsm wo Choose Your Assay Continued on next page 14 Calibrating the Qubit 2 0 Fluorometer Continued Running New Standards for Calibration continued If you have already performed a calibration for the selected assay Qubit 2 0 Fluorometer will prompt you to choose between reading new standards and using the previous calibration 100 200 300 400 500 Concentration ng mL Last Standards Created 19 Mar 2010 1 45 PM Read New Standards Note You can also press Standards to go to the Standards Screen from any other screen Press Yes to read new standards A prompt to insert Standard 1 appears on the screen 0 tI 200 200 400 S00 Concertinadion frigid Insert Standard Read Horne SEME Data Continued on next page 15 C
22. nsions Weight Dynamic range Processing time Light sources Excitation filters Emission filters Detectors Calibration type Tube type Warm up time Qubit 2 0 USB Drive 100 240 VAC 0 4A 50 60 Hz 9 VDC 1 33 A Indoor use only 15 40 C 20 80 non condensing Benchtop fluorometer 5 3 w x 8 5 1 x 1 6 h 13 3 cm x 21 3 cm x 4 cm oval shaped 11 5 oz 340 g 5 orders of magnitude lt 5 seconds sample Blue LED max 470 nm Red LED max 635 nm Blue 430 495 nm Red 600 645 nm Green 510 580 nm Red 665 720 nm Photodiodes measurement capability from 300 1 000 nm 2 or 3 point standard 0 5 mL Real Time PCR polypropylene tubes 2 seconds 2 Gigabyte Methods Getting Started Installing the Qubit 2 0 Fluorometer The Qubit 2 0 Fluorometer is a stand alone instrument that does not require a connection to a computer ie After unpacking the instrument place the instrument on a flat level dry surface Plug one end of the supplied power cord into the Qubit 2 0 Fluorometer To the other end attach the appropriate plug adaptor based on the electrical outlet configuration in your country Plug the power cord into the electrical outlet Be sure to use only the power cord supplied with your instrument Powering the instrument with an unapproved power cord may damage the instrument The instrument is automatically powered on when it is plugged in To
23. number of characters you can enter is 20 Note If you have selected to rename multiple lines at once files will be differentiated by numbers at the end of the name The numbers will appear in ascending order with the most recent sample starting with 1 e g SAMPLE_1 SAMPLE_ 2 etc Press OK to save the name you have entered as the file name and to go back to the Data Screen Continued on next page 26 Data Handling Continued Saving the Data to the Qubit 2 0 USB Drive The Qubit 2 0 Fluorometer is designed for stand alone use it does not require the use of an external computer However to archive data and generate reports you may transfer the numeric data stored in the CSV file to your computer using the USB drive and import the file into any spreadsheet program To archive your data 1 Insert the Qubit 2 0 USB drive into the USB port 2 Press Data to access the Data Screen 16 Aug 2010 16 Aug 2010 16 Aug 2010 16 Aug AHD i i Rename Home Standards Sample Note A green dot on the USB icon indicates that the instrument recognizes the USB drive a red dot indicates that the USB drive is not inserted into the USB port or that the instrument does not recognize the USB drive Continued on next page 2h Data Handling Continued Saving the Data to the Qubit 2 0 USB Drive continued 3 Select the data file to save by touching the corresponding line of the spreadsheet on the
24. ontact information visit our web site at www invitrogen com
25. roduce bubbles into samples Slight tapping on the tube wall or brief centrifugation will often help dissipate bubbles Continued on next page 30 Troubleshooting Continued Low Reading a 100 Am m0 400 Boo Concentration ngm ng ml Read Ment Sample Home Standards Your sample is out of range Use a sample that is more concentrated or use a lower dilution i e 20 uL in 180 pL instead of 10 uL in 190 pL View the Check Standards Screen to confirm that the values for the samples fall between the values of the standards see page 18 Ensure that you have prepared the Qubit working solution correctly 1 200 dilution using the buffer provided in the kit Ensure that you have prepared the standard tubes correctly 10 uL of each standard in 190 pL of Qubit working solution Ensure that the standard and sample tubes are filled to 200 uL Protect the Qubit reagent and working solutions from light Select the correct Qubit 2 0 Fluorometer assay for the Qubit assay you are performing and calibrate the fluorometer correctly Standards must be used in the correct order Ensure that the assay is performed entirely at room temperature Continued on next page 31 Troubleshooting Continued High Reading 12 Mac 2010 12 30 PM to 200 mo 400 S Concentration ingimL ng ml Read Ment Sample Home Standands e Your sample is out of range Use a sample that is less concentra
26. ront of the instrument is the user interface containing the buttons for all the functions needed and displays data from the assays Sample chamber is used to insert the assay tube containing the sample into the fluorometer for analysis USB port allows you to transfer and save data to your computer for record keeping and printing purposes The Qubit 2 0 USB drive supplied with the instrument or any other standard USB drive is inserted into the USB port for data transfer SE Power inlet Sample chamber Touch screen USB port Continued on next page Description of Qubit 2 0 Fluorometer Continued User Interface Use the touch screen of the Qubit 2 0 Fluorometer to operate the instrument The touch screen consists of e The touch screen buttons to operate the instrument including alphanumeric buttons to enter sample names e The digital display showing the data and graphs for the standards and sample O wma nam vo W Choose Your Assay Touch DNA screen button RNA Protein ad Dni BFE Digital Gonoeniration rg r display Lad Shonda it 19 Moa a a Pe Read New Standards Yes Alpha numeric buttons Home Standards Product Specifications Qubit 2 0 Fluorometer Specifications Environmental Conditions Operating power Frequency Electrical output Installation site Operating temperature Maximum relative humidity Instrument Specifications Instrument type Instrument dime
27. s Jan O10 Date 7 Feb 2011 8 2012 12 i Time l 32 PM 33 Set Cancel Home Standards Sample Data 2 Select the date and time by bringing the desired value to the center position on the roller wheel Press Set to make the date time changes The updated date time is displayed on the top of the window Note If you select Cancel your changes are not saved and the instrument returns to the screen that was last displayed To adjust the screen brightness press the Brightness button ES located on the top right of the touch screen and move the slider bar to the desired brightness level na Mac 2010 12 30 PM Choose Your Assay DNA RNA mm Protein a Standards Sample Continued on next page 11 Getting Started Continued Updating the To update your Qubit 2 0 Fluorometer with the latest firmware Firmware available from Invitrogen follow these steps 1 Download the latest firmware to your USB drive from our website www invitrogen com qubit 2 Press the Version button located on the top portion of the touch screen The Version Screen is displayed 19 ay 2010 12 30 PM v20 ae To update 1 Download new firmware fram our website www Invitrogen camyqubit to your USB drive 2 Insert your USB drive into the Qubit 3 Press Update button SE Update 3 Insert the USB drive into the USB port and press Update Note A green dot on the Update button indicates t
28. t temperature fluctuations within your laboratory you should determine the level of comfort you have using the calibration data stored from the last time the instrument was calibrated Remember also that the fluorescence signal in the tubes containing the standards and the samples is stable for not longer than 3 hours See Figure 1 in How the Qubit 2 0 Fluorometer Calculates Concentration page 33 for an example of the calibration curve used to generate the quantitation results 35 Accessory Products Qubit 2 0 Products The following products can be used with the Qubit 2 0 Fluorometer and are available separately from Invitrogen For more information visit www invitrogen com or contact Technical Support page 38 Product Quantity Catalog no Oubit 2 0 Fluorometer 1 each 032866 Oubit Quantitation Starter Kit 1 kit 032871 Qubit Quantitation Lab Starter Kit 1 kit 032872 Qubit 2 0 Fluorometer USB 1 each 032867 Qubit 2 0 Fluorometer International 1 each 032868 Power Cord replacement Qubit Assay Tubes 500 tubes 032856 Qubit RNA BR Assay Kit 100 assays Q10210 20 1 000 ng 500 assays Q10211 Qubit ssDNA Assay Kit 1 200 ng 100 assays Q10212 Qubit dsDNA BR Assay Kit 100 assays 032850 2 1 000 ng 500 assays 032853 Qubit dsDNA HS Assay Kit 100 assays Q32851 0 2 100 ng 500 assays 032854 Qubit RNA Assay Kit 5 100 ng 100 assays 032852 500 assays Q32855 Qubit Protein Assay Kit 0 25 5 pg 100 assays 03
29. ted e View the Check Standards Screen to confirm that the values for the samples fall between the values of the standards see page 18 e Ensure that the lid is closed while reading standards and samples e Prepare samples and standards according to the instructions in the Qubit assay kit you are using e Ensure that the assay is performed entirely at room temperature 32 Appendix Critical Qubit Assay Considerations How the Qubit 2 0 Fluorometer Calculates Concentration The Qubit 2 0 Fluorometer generates concentration data based on the relationship between the two Standards used in calibration three for the Qubit protein assay The plot below shows the line corresponding to the curve fitting algorithm a modified Hill plot used in the calculation of concentration data for the Qubit RNA assay For reference the positions of the Standards and a set of data points from an actual experiment are shown superimposed onto the line This plot demonstrates that the curve fitting algorithm gives accurate values for quantitation Relative fluorescence 0 100 200 300 400 500 600 Concentration of RNA ng mL Figure 1 The curve fitting algorithm used to determine concentration in the Qubit RNA assay Data for the Qubit dsDNA BR Qubit dsDNA HS Qubit ssDNA Qubit RNA BR and Qubit protein assays are generated by similar algorithms Continued on next page 33 Critical Qubit Assay Consi
30. ter or other material enters the instrument the adaptor or power inlet disconnect the power cord and contact a service person For operating environment refer to Product Specifications page 8 Do not touch the main plug or power cord with wet hands Always ensure that the power supply input voltage matches the voltage available in your location Do not install the instrument on a slant or a place prone to vibrations vibrations increase the risk of instrument malfunction or damage Plug the power cord firmly into the wall outlet and AC adapter To avoid potential shock hazard make sure that the power cord is properly grounded Be sure to position the equipment so that it is easy to disconnect the instrument Turn off the instrument before unplugging the power cord and or moving the instrument If the instrument is broken or dropped disconnect the power cord and contact a service person Do not disassemble the instrument Use only authorized accessories adaptor power cord and USB drive Introduction Description of Qubit 2 0 Fluorometer Qubit 2 0 Fluorometer Features The Qubit 2 0 Fluorometer is a benchtop fluorometer for the quantitation of DNA RNA and protein using the highly sensitive and accurate fluorescence based Qubit quantitation assays Use of the state of the art dyes selective for dsDNA RNA and protein minimizes the effects of contaminants in your sample that affect the quantitatio
31. utton on the bottom of the screen or Read Next Sample Note When you navigate away from the Dilution Calculator Screen the Qubit 2 0 Fluorometer saves the last values for the sample volume and the units in the Dilution Calculator Screen only Returning to the Dilution Calculator Screen displays these last selected values Continued on next page 23 Calculating the Original Sample Concentration Continued Saving the 1 Calculation to the Qubit 2 0 Fluorometer To save the data from your calculation to the Qubit 2 0 Fluorometer press Save on the Dilution Calculator Screen O amemo amm a OSHA HS Volume of Sample Used Saving calculation to Qubit ee Read Next Save Sampi vE one Standards The last calculated value of your measurement is saved in the CSV file and tagged with a time and date stamp G tg Mar 2010 12 30 PM Name Date 2215 pm 16 Ava 20 JASON 16 Aug 2010 JASON 2 16 Aug 2010 JASON 16 Aug 2000 ote 7 eran Hone Stendands Sample To review rename and save the data to the Qubit 2 0 USB drive see Data Handling next page 24 Data Handling Introduction The Qubit 2 0 Fluorometer presents comprehensive data with graphic reports and it allows saving of data as a CSV comma separated value file for sample comparisons Each measurement data point in the CSV file is numbered and exhibits a time and date stamp Reviewing the 1 To re
32. view the data saved on the Qubit 2 0 Fluorometer press Data Data on any screen The Data Screen displays the data in a spreadsheet format with additional data columns hidden to the right The most recent data point is displayed in the first line of the spreadsheet 2 30 15 pm 16 Aw 2010 JASON 3 16 Aug 2010 JASON 2 16 Aug 2010 JASON 1 16 Ag 2010 cote E Rename Hame Standards aa mple 2 Drag the scrollbars with a finger to move continuously across the spreadsheet to access the data columns hidden to the right C ta Mae 2000 12 30 PM Dilution Factor Std 1 RFU 6 75 75 i 6 35 638 tote Rename i Rename Hane Standands Sample one Standards Sample Continued on next page 25 Data Handling Continued Renaming Data 1 Files Select the data file to rename by touching the corresponding line of the spreadsheet on the Data Screen The selected line will be highlighted If no lines are selected you are prompted to select a line or multiple lines to rename Note To select multiple files highlight each line by one one or press the column headers to highlight all files To deselect all rows simply press the column header again Press Rename to enter a name for the selected line Alphabet Keys Screen is displayed To switch to the Numeric Keys Screen press 123 L Mar 2000 12 30 PM Standards Sample Home Standards Enter the file name using the keypad buttons displayed on the Save menu The maximum
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