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C M Y CM MY CY CMY KLightCycler® 480Gene Scanning SoftwareVersion 1.5www.roche-applied-science.com

10LightCycler® 480 Gene Scanning SoftwarePrologueSoftware License AgreementGeneral Information6 Licensee may not sublicense, assign or transfer the li

11PrologueSoftware License AgreementDuration and Termination8 The Agreement is effective until terminated. Licensee may terminate this Agreement at an

12LightCycler® 480 Gene Scanning SoftwarePrologueConventions Used in this ManualConventions Used in this ManualVII Text ConventionsTo impart informati

13PrologueWarnings and PrecautionsWarnings and PrecautionsVIII The LightCycler® 480 System is equipped with software, enabling the user of the Product

14LightCycler® 480 Gene Scanning SoftwareGene Scanning with the LightCycler® 480 System Gene Scanning with the A LightCycler® 480 System“Gene scann

15Gene Scanning with the LightCycler® 480 System High Resolution MeltingHigh Resolution Melting1 The key technique in gene scanning, High Resolution M

16LightCycler® 480 Gene Scanning SoftwareGene Scanning with the LightCycler® 480 System High Resolution MeltingThis figure shows how a High Resolution

17Gene Scanning with the LightCycler® 480 System Components of the LightCycler® 480 System Gene Scanning AssayComponents of the LightCycler2 ® 480 S

18LightCycler® 480 Gene Scanning SoftwareGene Scanning with the LightCycler® 480 System Components of the LightCycler® 480 System Gene Scanning Assay

19Gene Scanning with the LightCycler® 480 System Components of the LightCycler® 480 System Gene Scanning Assay Temperature shifting:3. The next step

20LightCycler® 480 Gene Scanning SoftwareGene Scanning with the LightCycler® 480 System Components of the LightCycler® 480 System Gene Scanning Assay

21Gene Scanning with the LightCycler® 480 System Advantages of the High Resolution Melting TechniqueAdvantages of the High Resolution 3 Melting Tech

22LightCycler® 480 Gene Scanning SoftwarePreparing a LightCycler® 480 System Gene Scanning ExperimentDesigning the Gene Scanning AssayPreparing a Li

23Preparing a LightCycler® 480 System Gene Scanning Experiment Sample MaterialSample Material2 Because a LightCycler® 480 System Gene Scanning experi

24LightCycler® 480 Gene Scanning SoftwarePreparing a LightCycler® 480 System Gene Scanning Experiment PCR PrimersPCR Primers3 Design PCR primers t

25Preparing a LightCycler® 480 System Gene Scanning Experiment PCR ReagentsPCR Reagents4 Hot-start PCR techniques are strongly recommended for

26LightCycler® 480 Gene Scanning SoftwarePreparing a LightCycler® 480 System Gene Scanning Experiment PCR ParametersPCR Parameters5 Data obtained fro

27Preparing a LightCycler® 480 System Gene Scanning Experiment PCR ParametersIn case you do not know the melting temperatures of your PCR

28LightCycler® 480 Gene Scanning SoftwarePreparing a LightCycler® 480 System Gene Scanning Experiment Rapid Detection of Amplification ArtifactsRapid

29Installing the LightCycler® 480 Gene Scanning SoftwareInstalling the LightCyclerC ® 480 Gene Scanning SoftwareThe LightCycler® 480 System Gene Scann

3Table of Contents Prologue 5I Revision History ...

30LightCycler® 480 Gene Scanning SoftwareInstalling the LightCycler® 480 Gene Scanning Software�The LightCycler® 480 Gene Scanning Software is install

31Performing Gene Scanning AnalysisPerforming Gene Scanning D AnalysisYou can perform a Gene Scanning analysis on any experiment that contains a

32LightCycler® 480 Gene Scanning SoftwarePerforming Gene Scanning Analysis �Click Sample Editor in the Module bar, then select the workflow Scanning. A

33Performing Gene Scanning Analysis �The Create new analysis dialog opens. Select an analysis subset and an experimental program from the Program list

34LightCycler® 480 Gene Scanning SoftwarePerforming Gene Scanning Analysis To perform a Gene Scanning analysis:�Using the Standards multi-select butto

35Performing Gene Scanning Analysis �Normalization ►Click the Normalization tab to normalize the melting curves. Fluorescence values acquired during

36LightCycler® 480 Gene Scanning SoftwarePerforming Gene Scanning Analysis �LightCycler® 480 Gene Scanning Software automatically places the temperatu

37Performing Gene Scanning Analysis �Temperature Shifting ►Click the Temperature Shift tab to reset the temperature axis (x-axis) of the melting curve

38LightCycler® 480 Gene Scanning SoftwarePerforming Gene Scanning Analysis �In the Results table, deselect any samples you do not want included in ana

39Performing Gene Scanning Analysis �Difference Plot ►Click the Difference Plot tab in the charts area to view the ► Normalized and Shifted Melting C

4LightCycler 480® Gene Scanning SoftwareTable of ContentsE Supplementary Functions 431 Gene Scanning Template ...

40LightCycler® 480 Gene Scanning SoftwarePerforming Gene Scanning Analysis To customize the Difference Plot display:�In the Difference Plot tab, click

41Performing Gene Scanning Analysis To rename groups:On the Groups tab in the Results table, you can change the name associated with each vari-ant/gen

42LightCycler® 480 Gene Scanning SoftwarePerforming Gene Scanning Analysis �Selecting a new genotype group from the New Call list and clicking Apply c

43Supplementary FunctionsGene Scanning TemplateSupplementary FunctionsE Gene Scanning Template1 A Gene Scanning analysis template contains the followi

44LightCycler® 480 Gene Scanning SoftwareSupplementary FunctionsResult Control Result Control 2 The LightCycler® 480 Gene Scanning Software uses a con

45Supplementary FunctionsUnlicensed FeaturesUnlicensed Features3 The following statements define the behavior of the LightCycler® 480 Software when a g

46LightCycler® 480 Gene Scanning SoftwareSupplementary FunctionsOrdering InformationOrdering Information4 Roche Applied Science offers a large selecti

47Supplementary FunctionsOrdering InformationPCR ReagentsLightCycler® 480 High Resolution Melting Master 5 × 100 µl (500 reactions, 20 µl each) 04 909

C M Y CM MY CY CMY Kwww.roche-applied-science.comPublished byRoche Diagnostics GmbHRoche Applied Science68298 MannheimGermany 2008 Roche Diagnostics G

5PrologueRevision HistoryPrologueRevision HistoryI Version Revision Date1.0 February 20072.0 February 2008© Copyright 2008, Roche Diagnostics GmbH. Al

6LightCycler® 480 Gene Scanning SoftwarePrologueTrademarksTrademarksIII LIGHTCYCLER, LC, MAGNA PURE and HIGH PURE are trademarks of Roche.Other brands

7PrologueSoftware License AgreementSoftware License AgreementVI Read the following terms and conditions of this Software License Agreement (“Ag

8LightCycler® 480 Gene Scanning SoftwarePrologueSoftware License AgreementLicensee may not:Use the Software, in whole or in part, except as expressly

9PrologueSoftware License AgreementDisclaimer of Warranties 4 THE WARRANTY SET FORTH IN THE PREVIOUS PARAGRAPH, IS IN LIEU OF ALL OTHER WARRANTIES, EX