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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.

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

Содержание Qubit 2.0

Страница 1: ...llen Way Carlsbad CA 92008 T 1 760 603 7200 F 1 760 602 6500 E tech_support invitrogen com For country specific contact information visit our web site at www invitrogen com Qubit 2 0 Fluorometer Catalog no Q32866 Rev Date 4 October 2010 MAN0003231 ...

Страница 2: ...d 9 Guidelines for Using the Qubit 2 0 Fluorometer 13 Calibrating the Qubit 2 0 Fluorometer 14 Reading Samples 19 Calculating the Original Sample Concentration 22 Cleaning and Maintenance 29 Troubleshooting 30 Appendix 33 Critical Qubit Assay Considerations 33 Accessory Products 36 Regulatory Requirements 37 Technical Support 38 Purchaser Notification 39 ...

Страница 3: ...the Instrument 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 Registering Your Instrument Visit probes invitrogen com qubit to register your instr...

Страница 4: ...nput 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 d...

Страница 5: ...accuracy even with very dilute samples Features 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 fluoresc...

Страница 6: ...in your country Touchscreen located in the front 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...

Страница 7: ...uch screen of the Qubit 2 0 Fluorometer to operate the instrument The touch screen consists of The touch screen buttons to operate the instrument including alphanumeric buttons to enter sample names The digital display showing the data and graphs for the standards and sample ...

Страница 8: ... fluorometer Instrument dimensions 5 3 w 8 5 l 1 6 h 13 3 cm 21 3 cm 4 cm oval shaped Weight 11 5 oz 340 g Dynamic range 5 orders of magnitude Processing time 5 seconds sample Light sources Blue LED max 470 nm Red LED max 635 nm Excitation filters Blue 430 495 nm Red 600 645 nm Emission filters Green 510 580 nm Red 665 720 nm Detectors Photodiodes measurement capability from 300 1 000 nm Calibrati...

Страница 9: ...into the Qubit 2 0 Fluorometer To the other end attach the appropriate plug adaptor based on the electrical outlet configuration in your country 3 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 4 The instrument is automatically powered on when it is plugg...

Страница 10: ...creen 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 Home Screen 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 th...

Страница 11: ...ct 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 Adjusting Screen Brightness To adjust the screen brightness press the Brightness button ...

Страница 12: ...reen The Version Screen is displayed 3 Insert the USB drive into the USB port and press Update Note A green dot on the Update button 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 After the successful update of the firmware the following screen is displayed befor...

Страница 13: ...e 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 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 ...

Страница 14: ...ation of the appropriate standard solutions For instructions on the preparation of these standards see the instructions that accompany the assay you are using Materials Needed 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 ...

Страница 15: ...bration for the selected assay Qubit 2 0 Fluorometer will prompt you to choose between reading new standards and using the previous calibration Note You can also press Standards to go to the Standards Screen from any other screen 3 Press Yes to read new standards A prompt to insert Standard 1 appears on the screen Continued on next page ...

Страница 16: ... 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 d...

Страница 17: ...fied by a large black circle 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 Using the Last Calibration You can choose to apply the previous calibration to your sample readings by pressing No when the Qubit 2 0 Fluorometer prompts you to choose between reading new standards and usin...

Страница 18: ... 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 much higher than that of Standard 2 and the reading of your sample should be no more tha...

Страница 19: ...ons on the preparation of your samples see the instructions that accompany the assay you are using Materials Needed Qubit assay kit you want to use to quantitate your samples see page 33 for available Qubit assay kits and ordering information 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 Qu...

Страница 20: ...ed 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 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 Con...

Страница 21: ...tandards Screen Pressing Show Graph brings the graph back onto the screen 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 ...

Страница 22: ...n Calculator 1 To calculate the concentration of your original sample press Calculate Stock Conc The Dilution Calculator Screen containing the volume roller wheel is displayed 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 meas...

Страница 23: ...he 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 μL the button displays pg μL 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 button ...

Страница 24: ... To save the data from your calculation to the Qubit 2 0 Fluorometer press Save on the Dilution Calculator Screen The last calculated value of your measurement is saved in the CSV file and tagged with a time and date stamp 2 To review rename and save the data to the Qubit 2 0 USB drive see Data Handling next page ...

Страница 25: ...its a time and date stamp Reviewing the Data 1 To review the data saved on the Qubit 2 0 Fluorometer press 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 Drag the scrollbars with a finger to move continuously across the spreadsheet to access ...

Страница 26: ...lumn header again 2 Press Rename to enter a name for the selected line Alphabet Keys Screen is displayed To switch to the Numeric Keys Screen press 123 3 Enter the file name using the keypad buttons displayed on the Save menu The maximum 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...

Страница 27: ... 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 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 instru...

Страница 28: ...tire CSV file will be saved to the USB drive if no lines are selected 4 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 5 To delete all data from the CSV file and start with a blank file press the Clear Data button 6 Transfer the Qubit 2 0 USB drive to the USB port on your PC You may open...

Страница 29: ...Fluorometer to avoid damaging the instrument Do not expose the Qubit 2 0 Fluorometer to direct sunlight Clean the surface of the Qubit 2 0 Fluorometer with a damp cloth To clean 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 damage...

Страница 30: ...e 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 ...

Страница 31: ...ion correctly 1 200 dilution using the buffer provided in the kit Ensure that you have prepared the standard tubes correctly 10 μL of each standard in 190 μL of Qubit working solution Ensure that the standard and sample tubes are filled to 200 μL 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 cali...

Страница 32: ...ds Screen to confirm that the values for the samples fall between the values of the standards see page 18 Ensure that the lid is closed while reading standards and samples Prepare samples and standards according to the instructions in the Qubit assay kit you are using Ensure that the assay is performed entirely at room temperature ...

Страница 33: ...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 Figure 1 The curve fitting algorithm used to determine concentration in the Qubit RNA assay ...

Страница 34: ...ms the solution and results in a low reading 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 Incubation Time 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 th...

Страница 35: ...the tube from the instrument and let it equilibrate to room temperature for 30 seconds before taking another reading Calibrating the Qubit 2 0 Fluorometer 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 y...

Страница 36: ...arter Kit 1 kit Q32872 Qubit 2 0 Fluorometer USB 1 each Q32867 Qubit 2 0 Fluorometer International Power Cord replacement 1 each Q32868 Qubit Assay Tubes 500 tubes Q32856 Qubit RNA BR Assay Kit 20 1 000 ng 100 assays 500 assays Q10210 Q10211 Qubit ssDNA Assay Kit 1 200 ng 100 assays Q10212 Qubit dsDNA BR Assay Kit 2 1 000 ng 100 assays 500 assays Q32850 Q32853 Qubit dsDNA HS Assay Kit 0 2 100 ng 1...

Страница 37: ...3 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 Commu...

Страница 38: ...technical assistance call write fax or email Additional international offices are listed on our website www invitrogen com Corporate Headquarters 5791 Van Allen Way Carlsbad CA 92008 USA Tel 1 760 603 7200 Tel Toll Free 1 800 955 6288 Fax 1 760 602 6500 E mail tech_support invitrogen com Japanese Headquarters LOOP X Bldg 6F 3 9 15 Kaigan Minato ku Tokyo 108 0022 Tel 81 3 5730 6509 Fax 81 3 5730 65...

Страница 39: ...oduct 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 Ou...

Страница 40: ... 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 location 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 inclu...

Страница 41: ...ual 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 contact information visit our web site at www invitrogen com ...

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