Vector NTI Advance 11 Quick Start Guide
Contents
1. NTI Advance 11 User s Manual for more information 4 Inthe dialog box select the desired primer design parameters Note that most of these parameters have default values based on typical PCR primers 5 Click on The results will appear under PCR Analysis in the Text Pane PCR analysis results lt n ry Figure 7 Designing PCR primers within a selected region To save the PCR analysis results as a separate object in the database 1 Right click on the PCR Analysis folder in the Text Pane 2 Select Save as Analysis Result The saved results will be listed under the Analysis Results object type in the Vector NTI Explorer Figure 8 ama Pore 7 Figure 8 PCR analysis results listed in Vector NTI Explorer 10 Clone Two Fragments with Clone2Seq Clone2Seq offers the easiest way to clone two fragments To use Clone2Seq 1 In the Molecule Viewer go to Cloning Clone2Seq or click on the CloneZSeq button illoa the main toolbar To load Insert and or Vector click on Load Molecule Select fragment by Restriction Site by clicking on Site 1 shift clicking on Site 2 on each molecule Make sure the left terminus of the first fragment is compatible with the right terminus of the second fragment and the right terminus of the first fragment is compatible with the left terminus of the second fragment Add selected fragments by clicking on Add Fragment Clic2. describes Information concerning products not manufactured or distributed by Invitrogen Corporation is provided without warranty or representation of any kind and neither Invitrogen Corporation nor its affiliates will be liable for any damages Table of Contents Introduction Opening Vector NTI Advance 11 Local Database Database Backup Restore Molecule Viewer Selecting and Editing Molecule Sequences Designing PCR Primers from a Sequence Clone Two Fragments with Clone2Seq In Silico Gene Synthesis with ReGENeratoi Identifying Open Reading Frames ORFs Creating a Restriction Map Aligi Contig Assembly Add g Molecules nal Information Introduction This Quick Start Guide is designed to get you started using Vector NTI Advance 11 It provides brief descriptions of the Vector NTI Advance 11 graphical user interface including Vector NTI Explorer and the Molecule Viewer and step by step instructions for using the most common features and functions of the software The topics covered include locating the desired tools displaying molecules designing PCR primers cloning two fragments gene synthesis aligning molecules performing a restriction analysis and assembling contigs This guide assumes that you have a working knowledge of basic Microsoft Windows and Mac OS features and functions how to open and save files how to use your mouse and so on and that Vector NTI Adva
3. of two or more DNA RNA molecules The tool for doing this is called AlignX This tool can be launched from either the Molecule wer or Vector NTI Explorer To align sequences using Vector NTI Explorer 1 In the Explorer select the molecules that you want to align using SHIFT CLICK or CTRL key commands Figure 13 Go the Align menu and select AlignX Align Selected Molecules The AlignX Window will open with the molecules you selected listed in the upper left Text Pane In the AlignX Window use SHIFT CLICK or CTRL CLICK key commands to select two or more molecules in the Text Pane list to align To begin the alignment click on the Align button on the toolbar The alignment may take several minutes depending on the length and number of the molecules selected When the alignment is complete the results are displayed in the AlignX Window as shown in Figure 13 The AlignX Window has panes showing different similarity graphs and the points at which the sequences align natn p ro Akere nd E 8 EE Fi si Complexity and similarity plots Relatedness tee Figure 13 ing molecules 15 Contig Assembly Vector NTI Advance 11 can be used to assemble DNA fragments both text sequences and chromatograms from automated sequencers into longer contiguo
4. open reading frames ORFs in it based on start and stop codons within the molecule With DNA or RNA molecule open in the Molecule Viewer 1 Goto the Analyses menu and select ORF Figure 11 2 1 dialog box select the parameters for identifying and marking ORFs in the molecule 3 When you click on OK the sequences identified as ORFs will be marked with directional arrows in the Graphics Pane and Sequence Pane and the ORFs will be listed in the Text Pane 4 To identify an ORF in the different panes Click on a directional ORF arrow in the Graphics Pane to highlight its sequence in the Sequence Pane or Open a folder under Open Reading Frames in the Text Pane right click on the ORF name and select Find ORF to highlight it in the Graphics and Sequence Panes 5 To save an ORF to the feature map of the molecule right click on the ORF arrow in the Graphics Pane or the ORF folder in the Text Pane and select Add ORF to FMap Solr mp ORF marked in Graphic pane ow yo o meton ed Figure 11 Identifying ORFs 13 Creating a Restriction Vector NTI Advance 11 can analyze a DNA RNA molecule and identify the restriction sites in it using the software s comprehensive library of restriction enzymes With DNA or RNA molecule open in the Mol
5. Vector NTI Advance 11 Quick Start Guide Catalog no 12605050 12605099 12605103 Version 11 0 December 15 2008 12605022 Published by Invitrogen Corporation 5791 Van Allen Way Carlsbad CA 92008 USA www invitrogen com Copyright 2008 Invitrogen Corporation All rights reserved This document contains proprietary information of Invitrogen Corporation No part of this document including design cover design and icons may be reproduced or transmitted in any form by any means electronic photocopying recording or otherwise without prior written agreement from Invitrogen Corporation The software described in this document is furnished under a license agreement Invitrogen Corporation and its licensors retain all ownership rights to the software programs offered by Invitrogen Corporation and related documentation Use of the software and related documentation is governed by the license agreement accompanying the software and applicable copyright law Vector NTI Advance and Gateway are trademarks of Invitrogen Corporation in the United States and other countries Logos of Invitrogen Corporation are also trademarks registered in the United States and may be registered in other countries Other product and brand names are trademarks of their respective owners Generated in the United States Invitrogen Corporation reserves the right to make and have made changes without notice both to this publication and to the product it
6. agging in Graphics Pane 2 dragging in Sequence Pane To copy a molecule sequence 1 Select it as described above 2 Tocopyitto the Windows clipboard use the CTRL C keyboard command To copy the sequence as a text file go to the Edit menu and select Copy to gt File You will be prompted to select a format and enter a name for the file To delete a molecule sequence 1 Select it as described above 2 Click on the DELETE key on your keyboard To paste a molecule sequence 4 With the sequence in text format on the Windows clipboard click on the point in the Sequence Pane where you want to add the insert Click on CTRL V on your keyboard log will open displaying the sequence to be inserted 3 The Insert Sequence Click on OK to complete the insertion Designing PCR Primers from a Sequence Vector NTI Advance 11 can analyze a selected sequence and design PCR primers for it based parameters such as desired melting temperature Tm GC content and amplicon length With a DNA or RNA molecule open in the Molecule Viewer 1 Select the part of the sequence for which you want to design primers as described on the previous page Go to the Primer Design menu 3 Select Find PCR Primers to find primers within the sequence Figure 7 Select Amplify Selection to find primers in the regions before and after the sequence other amplification selections are available see the Vector
7. different types of molecular biology objects Each database record includes all the information for that object e g DNA molecule record includes the DNA sequence defined features of the molecule and other information Objects in the database can include molecules analysis results BLAST search results citations and other types of information Importcreate molecules Display analyze molecules Import analysis results Export molecules Import BLAST results Launch tools st of database records Database object type Figure 2 Vector NTI Explorer Local Database window Database objects in Vector NTI Advance are categorized by type DNA molecules protein molecules and so on Some molecules are installed with the software When you first open the software DNA RNA Molecules is the selected object type Click on the tab in the lower left comer of the Vector NTI Explorer to select from the other available database objects To open an object from the local database double click on the object name in the right hand pane of the Explorer Depending on the object type information about that object may be displayed in a dialog box or the object may be loaded into a viewer For example DNA RNA and protein molecules are displayed in the Molecule Viewer When you install Vector NTI Advance the default local database is created in a Note folder called VNTI Database in the root directory of y
8. ecule Viewer 1 Goto the Analyses menu and select Restriction Analyses gt Restriction Sites Figure 12 2 Inthe Restriction Map Setup dialog review the list of restriction enzymes in the Use Enzymes field These are the enzymes that will be used to identify the restriction sites Click on the lt Add gt Remove and gt gt Remove Alll buttons to add and remove enzymes from the list Note If you click on lt Add the Choose Database Enzymes dialog will open listing all the enzymes in the database Select enzymes in the list by clicking on them or click on the Select All button and then click on the OK button to add them to the Restriction Map Setup dialog 3 OK in the Restriction Map Setup dialog The restriction enzymes and their binding sites will be shown in the Graphics Pane and Sequence Pane The specific cut site of each enzyme will be the Text Pane I n o 9 ide Tk aril m ie T x Sere oe Stewed bein Rs i 3 M 068 DA zA Rest site shown it nen Graphics Pane i ien Rest site shown in Sequence Pane Figure 12 Creating a Restriction Map 4 Aligning Molecules Vector NTI Advance 11 can align the sequences
9. k on Clone Figure 9 Clone 2 fragments with Clone2Seq 11 In Silico Gene Synthesis with ReGENerator ReGENerator offers the fastest way to build your desired DNA from the ground up optimized for expression with any amino acid mutation you want and with the flanking sequences you need for expression purification or detection Then right from your desktop send your DNA sequence to our partner Blue Heron Bio s secure server for rapid synthesis To design the DNA from an amino acid sequence 1 With the source protein molecule loaded in the Molecule Viewer select Cloning ReGENerator or cick on the icon on the main Mutate the protein molecule by inserting deleting or replacing single or multiple amino acids Select the desired Codon Usage Table for the organism Select the desired Genetic Code RON You may also add attachment sequences to the terminus of the DNA Choose terminus 5 or 3 Choose Attachment 6 the back ranslated DNA click View Molecule 7 To send the DNA sequence for synthesis at Blue Heron click Send for Synthesis ich lo In Sec DNA Sequence prep me und Mahis Dusk Type EC e Emm Figure 10 In Silico Gene Synthesis with ReGENerator 12 Identifying Open Reading Frames ORFs Vector NTI Advance 11 can analyze a DNA RNA molecule and identify the
10. nce 11 is installed on your computer Opening Vector NTI Advance 11 The QuickStart Page is a single page that consolidates most commonly used modules tools and utilities that Vector NTI Advance provides To launch the QuickStart Page select Start gt All Programs gt Invitrogen gt Vector NTI Advance 11 gt Quick Start Vector NTI Advance 11 QuickStart Page Imm pem m p aratan Figure 1 QuickStart Page You can configure the software to open both the Molecule Viewer and Vector NTI Explorer when you select Vector NTI from the Start menu 1 Inthe Molecule Viewer window go to the Edit menu and select Options 2 Inthe General tab of the dialog select the Open Local Explorer At Startup checkbox 3 Click OK to make the change Local Database Vector NTI Explorer is the main tool for accessing the information in your local Vector NTI Advance database Using the Explorer you can import open export and organize molecules and other database items and launch other Vector NTI Advance modules Figure 2 To launch Vector NTI Explorer On QuickStart Page click on Local Database Inthe Molecule Viewer click on the Local Database icon From the Windows Start menu select Programs Invitrogen Vector NTI Advance 11 Vector NTI Explorer The local database in Vector NTI Advance contains records for
11. our computer e g Database Database Backup Restore Itis strongly recommended that the local database be backed up routinely You may launch the Database Backup manually or use the Database Backup Reminder to trigger the task automatically as configured Figure 3 To manually perform Database Backup To configure the Database Backup Reminder 4 To set a specific date or interval Figure 3 To restore a database From Vector NTI Explorer menu re Figure 4 From the Vector NTI Explorer menu select Database Backup Database Now From the Vector NTI Explorer menu select Database Database Backup Reminder e g backup every 15 days click on Set Reminder ET s Quran rom Diya 3 EN Database Backup select Database Database Restore Database Restore Molecule Viewer The Molecule Viewer displays information about DNA RNA and protein molecules To launch the Molecule Viewer Click on Molecule Viewer on the QuickStart page or From the Windows Start menu select Programs Invitrogen Vector NTI Advance 11 Vector NTI or Double click on a molecule name in the Vector NTI Explorer E open a molecule from within the Molecule Viewer click on the Open button 7 on the main toolbar and select the molecule name from the dialog box The molec
12. ragments in the assembly The Text Pane on the left lists the fragments in the assembly 9 Ifyou wish to edit the contig enable the Enhanced Edit Mode by clicking the icon Use Enhanced Edit Mode far left on the toolbar in the Contig window before making any reasonable changes 10 There are three trimming options in ContigExpress Fragments can be trimmed for ambiguities Phred quality scores and vector contamination Refer to the Vector NTI Advance 11 User s Manual for details Continued on the following page 16 Contig Assembly continued 4 er ae te History Viem 12 egrets m projet i Hl M Fragment Viewer i i i H u Figure 14 Assembling a Contig Additional Information Invitrogen s free technical support for Vector NTI Advance 11 is available exclusively through the web For more information check out the Software Support section of the Vector NTI website at http www invitrogen com VectorNTI To obtain personalized technical support by telephone or email you must have an annual support contract You may purchase an Advanced Support Contract by contacting Invitrogen at bioinfosales invitrogen com To receive technical support use the follo
13. ule will be loaded into the Molecule Viewer Pane specife E Tex Pane info about features ses ele 4 Graphics Pane Sequence Pane 6 wer window for a DNA molecule The Molecule Viewer window has different panes for displaying different types of information about the molecule as shown in Figure 5 Click inside a pane to make it the active pane The available tools and right click menu options will change depending on which pane is active igure 5 Molecule Use tools on the dropdown menus and toolbars to add information about the molecule and perform various analysis functions as described in the step by step instructions on the following pages Multiple molecules can be displayed in separate windows of the Molecule Viewer Selecting and Editing Molecule Sequences In the Molecule Viewer you can select part of a molecule sequence in several different ways Hold down the mouse button and drag the cursor across the sequence the Sequence Pane or Graphics Pane Figure 6 Go to the Edit menu select Set Selection and enter the sequence base pair range in the dialog box Click on a defined feature in the Graphics Pane Click on a defined feature in the Text Pane and click on Find 84 on the main toolbar The selected sequence will appear highlighted in both the Graphics Pane and the Sequence Pane Figure 6 Selecting a DNA sequence by 1 dr
14. us sequences or contigs The tool for doing this is called ContigExpress In Vector NTI Explorer or the Molecule Viewer 1 Goto the Assemble menu and select ContigExpress Open New Assembly Project Figure 144 2 In the ContigExpress Project Explorer go to the Project menu and select Add Fragments gt Select your fragment file type from the submenu list The Import Sequence dialog will open 3 Inthe Import Sequence dialog navigate to the directory containing your fragment sequence files Select the files and click on Open 4 Depending on the file type you may be prompted to list the fragments by their Windows file names or by their internal fragment names Select the desired option The fragments will be loaded in the ContigExpress Project Explorer 5 view a particular fragment double click on it in the Project Explorer list It will be loaded into the Fragment Viewer 6 When you are ready to perform contig assembly select the fragments in the ContigExpress Project Explorer 7 Click the Assemble Selected Fragments icon on the main toolbar Fragments will be analyzed and assembled into one or more contigs which will be listed in the Project Viewer along with the fragments in each contig 8 Double click on a contig in the list It wll be displayed in the Contig Viewer The Sequence Pane at the bottom shows the sequence of the assembly The Graphics Pane on the right shows the orientations of the f
15. wing contacts United States Phone 800 955 6288 x 67990 Toll free U S E mail bioinfosupport invitrogen com Europe Middle East Africa Asian Pacific Phone 44 781 696 2707 Email bioinfosupport invitrogen com 18
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