qPCR Instrument Calibration Kit
Contents
1. have been processed Page 3 Guidelines for Calibrating ABI PRISM Machines If you are using a different real time qPCR system than the models described below refer to the system s instrument manual for calibration guidelines Loading Volume Real Time qPCR Instrument Format Volume well ABI PRISM 7000 96 well plate 50 ul well ABI PRISM 7500 96 well plate 50 pl well ABI PRISM 7700 96 well plate 50 ul well ABI PRISM 7900 HT 96 well plate 50 pl well 384 well plate 20 ul well Plate Configurations ABI PRISM 7000 and 7500 Plate Configuration For the ABI PRISM 7000 and 7500 models each calibration dye needs to be aliquoted into every well of a 96 well plate for a total of 8 separate calibration plates ABI PRISM 7700 and ABI PRISM 7900 96 Well Plate Configuration 5 6 7 8 9 10 11 12 ABI PRISM 7900 HT 384 Well Plate Configuration 12 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 OZzZzr FxAcC TATAMIODS Page 4 Additional Products Product Amount Catalog no Custom Designed LUX Fluorogenic Primers www invitrogen com lux Platinum Quantitative PCR SuperMix UDG 100 rxns 11730 017 SuperScript III Platinum Two Step qRT PCR Kit 100 rxns 11734 050 SuperScript III Platinum One Step qRT PCR Kit 100 rxns 11732 020 Purchaser Notification Limited Use Label License No 1542. 3 Invitrogen life technologies qPCR Instrument Calibration Kit Cat No 11742 100 Store at 20 C in the dark Components 8 tubes Description The qPCR Instrument Calibration Kit provides qualified reagents for calibrating your real time qPCR system The kit includes common dyes used in real time qPCR SYBR Green I ROX TAMRA and dyes for use with LUX Fluorogenic Primers FAM JOE HEX TET and AlexaFluor 546 Real time qPCR instruments should be calibrated as part of regular instrument maintenance and prior to using new dyes for the first time when performing qPCR Refer to your real time qPCR instrument s user manual for specific instructions on calibration Components The qPCR Instrument Calibration Kit contains one tube of each of 8 calibration dyes in amber tubes Dye Volume Concentration FAM Calibration Dye 0 6 ml 10X SYBR Green Calibration Dye 0 6 ml 10X JOE Calibration Dye 0 6 ml 10X HEX Calibration Dye 0 6 ml 10X TET Calibration Dye 0 6 ml 10X AlexaFluor 546 Calibration Dye 0 6 ml 10X TAMRA Calibration Dye 0 6 ml 10X ROX Calibration Dye 0 6 ml 10X Instrument Compatibility This calibration kit is for use with real time qPCR instruments that have the option of being calibrated manually The calibration kit is compatible with but not limited to the following real time qPCR instruments ABI PRISM 7000 ABI PRISM 7500 ABI PRISM 7700 ABI PRISM 7900 HT Storage Store 10X or
3. SYBR Technology The manufacture use sale or import of this product may be subject to one or more of US patent Nos 5 436 134 5 534 416 5 545 535 5 658 751 5 863 753 6 664 047 pending applications and corresponding foreign equivalents owned by Invitrogen Corp The purchase of this product conveys to the buyer the non transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer whether the buyer is an academic or for profit entity The buyer cannot sell or otherwise transfer a this product b its components or c materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes The buyer may transfer information or materials made through the use of this product to a scientific collaborator provided that such transfer is not for any Commercial Purpose and that such collaborator agrees in writing a to not transfer such materials to any third party and b to use such transferred materials and or information solely for research and not for Commercial Purposes Commercial Purposes means any activity by a party for consideration and may include but is not limited to 1 use of the product or its components in manufacturing 2 use of the product or its components to provide a service information or data 3 use of the product or its components for the
4. ct nor any of its components was used in the manufacture of such product If the purchaser is not willing to accept the limitations of this limited use statement Invitrogen is willing to accept return of the product with a full refund For information on purchasing a license to this product for purposes other than research contact Molecular Probes Inc Business Development 29851 Willow Creek Road Eugene OR 97402 Tel 541 465 8300 Fax 541 335 0354 ABI PRISM is a registered trademark of the Applera Corporation 2005 Invitrogen Corporation All rights reserved For research use only Not intended for any animal or human therapeutic or diagnostic use
5. diluted calibration dyes at 20 C in the dark When thawing and using calibration dyes protect from direct light For most instruments calibration is recommended every 3 6 months Save the diluted dyes in the plates tubes used to do the calibration at 20 C in the dark until next calibration Calibration dyes may be reused for at least one year when stored correctly Caution Wear powder free gloves when handling the calibration dyes Contact Technical Service to obtain the MSDS for further precautions on kit components Quality Control Calibration dyes are tested first by measuring the emission peaks and second by analyzing the raw spectra data on an ABI PRISM 7700 Part no 11742 pps Rev Date 23 Feb 2005 This product is distributed for laboratory research use only CAUTION Not for diagnostic use The safety and efficacy of this product in diagnostic or other clinical uses has not been established For technical questions about this product call the Invitrogen TECH LINESM 800 955 6288 Form No 18057N Printed on Recycled Paper Page 2 General Protocol The following protocol describes the steps involved in calibrating real time qPCR instruments For most systems calibration should occur at initial setup before the use of new dyes and after maintenance service from the manufacturer For more detailed instructions refer to the manual for your particular instrument If you are using an ABI PRISM real time qPCR system re
6. fer to the next page for more details Important If you have previously calibrated your real time instrument for a dye included with this kit and wish to avoid overwriting the calibration setting you should give the new calibration setting a different name for example Invitrogen ROX Protocol 1 Thaw the 10X calibration dyes at room temperature away from direct light 2 Dilute each 10X calibration dye 1 10 in 1X TE for a total volume of 5 ml in a new tube 3 Aliquot the diluted calibration dyes into tubes or plates according to the suggested configuration and format for your real time qPCR system If you are using an ABI PRISM instrument see the guidelines on the next page Return any unused 1X calibration dyes to 20 C in light safe tubes 4 Centrifuge the diluted calibration dyes in tubes or plates for 2 minutes at low speed to ensure that bubbles are removed 5 Place diluted calibration tubes or plates into the sample tray of your real time qPCR machine 6 Runa pure dye spectral calibration on your real time qPCR system using the protocol described in the instrument s user manual 7 Analyze the data after completing the calibration run using the protocol described in the instrument s user manual 8 After processing wrap the diluted calibration dyes in aluminum foil to protect from light and return to 20 C If you are calibrating each dye with an individual plate return to step 5 and repeat until all plates
7. rapeutic diagnostic or prophylactic purposes or 4 resale of the product or its components whether or not such product or its components are resold for use in research Invitrogen Corporation will not assert a claim against the buyer of infringement of the above patents based upon the manufacture use or sale of a therapeutic clinical diagnostic vaccine or prophylactic product developed in research by the buyer in which this product or its components was employed provided that neither this product nor any of its components was used in the manufacture of such product If the purchaser is not willing to accept the limitations of this limited use statement Invitrogen is willing to accept return of the product with a full refund For information on purchasing a license to this product for purposes other than research contact Molecular Probes Inc Business Development 29851 Willow Creek Road Eugene OR 97402 Tel 541 465 8300 Fax 541 335 0504 Limited Use Label License No 183 AlexaFluor Dyes The manufacture use sale or import of this product may be subject to one or more of U S Patent Nos 5 696 157 6 130 101 6 716 979 pending applications and corresponding foreign equivalents owned by Invitrogen Corp The purchase of this product conveys to the buyer the non transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer whether the buyer is an academic or for profit en
8. tity The buyer cannot sell or otherwise transfer a this product b its components or c materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes The buyer may transfer information or materials made through the use of this product to a scientific collaborator provided that such transfer is not for any Commercial Purpose and that such collaborator agrees in writing a to not transfer such materials to any third party and b to use such transferred materials and or information solely for research and not for Commercial Purposes Commercial Purposes means any activity by a party for consideration and may include but is not limited to 1 use of the product or its components in manufacturing 2 use of the product or its components to provide a service information or data 3 use of the product or its components for therapeutic diagnostic or prophylactic purposes or 4 resale of the product or its components whether or not such product or its components are resold for use in research Invitrogen Corporation will not assert a claim against the buyer of infringement of the above patents based upon the manufacture use or sale of a therapeutic clinical diagnostic vaccine or prophylactic product developed in research by the buyer in which this product or its components was employed provided that neither this produ
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