Generating ABI PRISM® 7700 SDS Standard Curve Plots in a
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1. Quantity 1 00E 03 1 00E 03 1 00E 03 1 00E 03 2 00E 03 2 00E 03 Standard Curve 2 00E 03 2 00E 03 5 00E 03 5 00E 03 5 00E 03 5 00E 03 1 00E 04 4 00E 04 1 00E 04 1 00E 304 2 00E 404 200E 405 20 30 200E 406 43 2281 Log Quantity 2 00E 407 Applied A Biosystems Formatting the Graph Formatting the X and Y axis 1 To adjust the X axis and Y axis to the appropriate scale move the cursor over the appropriate axis and double click Change the scale accordingly Format Axis LN NI 2 Click OK N Microsoft Excel Exceldata xls Quantity 1 00E 403 1 00E 403 1 00E 403 1 00E 403 2 00E 403 3 2 00E 403 Standard Curve 2 00E 403 2 00E 403 5 00E 03 5 00E 403 5 00E 403 5 00E 403 1 00E 04 1 00E 404 1 00E 404 1 0DE 404 2 00E 404 EE 2 00E 405 2 00E 06 3 Log Quantity 2 00E 407 Applied A Biosystems Adding a Trend Line 1 To adda Trend Line place the cursor on a point of the graph and click once to highlight the data points N Microsoft Excel Exceldata xls P 5 r ujeeme amp s 00 4 0 Ez a p o7 D so Quantity 1 00E 403 1 00E 403 1 00E 403 1 00E 403 2 00E 403 33 07 Standar2. Paste the Quantities and Ct values from this data into a new Worksheet Create a column between the Quantities column and the Ct column gt I ue a N The Quantities need to be converted to their log value To do this click on the fx icon on the Toolbar Click on Math and Trig and select Log iN Applied RAS Blobystems 7 Convert the quantities into their log values in the appropriate column The log values will be displayed on the x axis and the Ct values on the y axis N Microsoft Excel Booki Quantity 1 00E 03 1 00E 403 1 00E 403 1 00E 403 2 00E 403 2 00E 403 2 00E 403 2 00E 403 5 00E 403 5 00E 403 5 00E 403 5 00E 403 1 00E 404 1 00E 404 1 00E 404 1 00E 404 2 00E 04 2 00E 05 2 00E 06 2 00E 07 Opening Chart Wizard 1 Go to the Chart Wizard icon located on the Excel tool bar and click to open N Microsoft Excel Booki AB Bibsysic Quantity 1 00E 403 1 00E 403 1 00E 403 1 00E 403 2 00E 403 2 00E 403 2 00E 403 2 00E 403 5 00E 403 5 00E 403 5 00E 403 5 00E 403 1 00E 404 1 00E 404 1 00E 404 1 00E 404 2 00E 404 2 00E 405 2 00E 06 2 00E 407 ystems Bio 1g LI Log Quantity Ct 3 0 3 0 3 0 3 0 33 33 3 3 Chart Wizard Icon La Log Quantity Ct 3 0 Chart Wizard Step 1 Chart Type 1 For the chart type select XY scatter and the chart sub type highlighted Chart Wizard Step 1 of 4 Chart Type 2 Click Next Chart Wizard Step 2 Cha
3. in diagnostic procedures Information in this document is subject to change without notice Applied Biosystems assumes no responsibility for any errors that may appear in this document This document is believed to be complete and accurate at the time of publication In no event shall Applied Biosystems be liable for incidental special multiple or consequential damages in connection with or arising from the use of this document Applied Biosystems and ABI PRISM are registered trademarks of Applera Corporation or its subsidiaries in the US and certain other countries TaqMan is a registered trademark of Roche Molecular Systems Inc Macintosh is a registered trademark of Apple Computer Inc All other trademarks are the sole property of their respective owners 117GU14 01 Part Number 4370924 Revision A iN Applied RAS Biosystems
4. Generating ABI Prism 7700 Standard Curve Plots in a Spreadsheet Program Overview The goal of this tutorial is to demonstrate the procedure through which analyzed data generated within an ABI PRISM 7700 data file can be exported and regenerated in Microsoft Excel This tutorial demonstrates how to generate a Standard Curve The following tutorial should be used as a guideline only For further information on how to use Microsoft Excel and Chart Wizard please refer to the Microsoft Excel user manual Applied Biosystems does not support Microsoft Excel or any other Microsoft Software Manipulations within the ABI PRISM 7700 software 1 Open the SDS data file to be analyzed 2 Analyze the data and set the baseline and threshold according to Applied Biosystems guidelines To obtain more information on baseline and threshold setting guidelines please refer to the ABI Prism 7700 Setting Baselines and Thresholds tutorial available on line at http www appliedbiosystems com support apptech 3 From the File menu select Export and Results You may either export all of the data or data from selected wells click the appropriate button 4 The default file name will be data results Provide a name and file location for this data It is suggested that you keep the suffix results Manipulations within Microsoft Excel 1 Opena new Workbook in Microsoft Excel Select File Open Open the results file Copy and
5. d Curve 2 00E 403 2 00E 403 2 00E 403 amp 00E 03 5 00E 403 5 00E 403 S 00E 03 J 46 Series Ct Point 3 7 4 00E 04 40 2371 3 7 24 98 3 5 Log Quantity 2 From the Chart tab on the menu bar select Add Trendline Add Trendline Ei 7 Ej Jil hu I 3 Under the Type tab highlight Linear Add Trendline a aea 4 Click OK Applied A Biosystems Displaying Equation and R Squared Values 1 Under the Options tab select the Display equation on chart and Display R Squared value on chart boxes Add Trendline 2 Click OK 3 The equation will be displayed on the chart which can be moved by clicking and dragging it to a new location N Microsoft Excel Exceldata xls Quantity 1 00E 403 1 00E 403 1 00E 403 1 00E 403 2 00E 403 3 Standard Curve y 3 5115x 37 678 200E 403 R 0 995 2 00E 403 2 00E 403 5 00E 403 5 00E 403 5 00E 403 l 5 00E 03 Linear Ct 1 00E 04 MI Linear Ct 1 00E 404 1 00E 404 1 00E 04 j 2 00E 04 2 00E 05 3 5 2 00E 06 Log Quantity 2 00E 407 Applied A Biosystems 2002 Applied Biosystems All rights reserved For Research Use Only Not for use
6. rt Source Data 1 Chart Wizard now asks for a Data Range Chart Wizard Step 2 of 4 Chart Source Data Applied A Biosystems 2 To select a data range place your cursor at the top corner of the data points to be used Click and drag to highlight the data points Include the data for both axes The data points will be plotted in the Chart Wizard Quantity ORT Se Ee Ee 2 D 21 100E 03 Chart Wizard Step 2 of 4 Chart Source Data Data range X D 2 HET 1 00E 403 Sheet D 4 424 1 00E 403 1 00E 03 2 00E 03 2 00E 03 2 00E 03 2 00E 03 5 00E 403 5 00E 03 5 00E 403 5 00E 403 1 00E 04 1 00E 404 1 00E 04 1 00E 04 2 00E 04 2 00E 05 2 00E 06 2 00E 07 Source Data 3 Click Next Applied i A Biosystems Chart Wizard Step 3 Chart Options 1 Click on the Titles tab and name the chart title and axes Chart Wizard Step 3 of 4 Chart Options Standard Curve Standard Curve Log Quantity ct values Ct Walmez 2 0 3 0 Log Quantity 2 Click Next Chart Wizard Step 4 Chart Location 1 Select a location for the chart The chart can be created in a new sheet or within the current spreadsheet In this example the chart will be created in the open spreadsheet Chart Wizard Step 4 of 4 Chart Location H 100 1
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