Introduction to VectorNTI September 14, 2011 Ansuman Chattopadhyay, PhD, Head Molecular Biology Information Service Health Sciences Library System University of Pittsburgh ansuman@pitt.edu http://www.hsls.pitt.edu/guides/genetics Topics Vector NTI Advance 11.0 Suite Overview How to Import or Create New Sequences Sequence Alignments: AlignX Sequence Assembly: ContigExpress Analyze Sequences: PCR and Primers, Restrictions and Mutagenesis How to create Vector Maps Video Tutorials • VectorNTI user inter phase : a short walk-through http://media.hsls.pitt.edu/media/clres2705/vnti_walkthrough_part1.swf (16 min) http://media.hsls.pitt.edu/media/clres2705/vnti_walkthrough_part2.swf (10 min) • Import your DNA of interest into VectorNTI http://media.hsls.pitt.edu/media/clres2705/vnti_inportdna.swf (14 min ) • Create a multiple protein sequences alignment plot http://media.hsls.pitt.edu/media/clres2705/vnti_msa.swf (17 min) • Design PCR primers using VectorNTI http://media.hsls.pitt.edu/media/clres2705/vnti_pcr.swf (11 min) • Clone a fragment from pBR322 into pUC19 (in sillico molecule construction) http://media.hsls.pitt.edu/media/clres2705/vnti_cloning.swf (15 min) Exercise PLCg1: Mouse: Rat: Cow: Dog: Zebra fish: NP_067255 NP_037319 NP_776850 XP_542998 NP_919388 Human: NP_002651 NP_067255,NP_037319,NP_776850,XP_542998,NP_919388,NP_002651 Important Links PowerPoint Guide to Molecule Construction and Design PowerPoint Guide to Managing the Vector NTI Database Northwestern University VectorNTI tutorial A Molecule Construction Example Task – Clone a fragment from pBR322 into pUC19 ☼ Donor fragment: pBR322, 5’EcoRI—3’AvaI ☼ Recipient fragment: pUC19, 5’SmaI—3’EcoRI Getting started 1. Open pBR322 and pUC19 in Vector NTI 2. Arrange the display window to display 2 molecules on the same screen: ☺ Select Menu > Window > Tile Vertical Hands - On Epidermal Growth Factor receptor is considered as one of the bio-markers for several forms of cancer including Breast and Ovarian cancers. Create a Vector NTI file for the gene EGFR (Accession Number: P00533 ) Design PCR primers to amplify any one domain of your choice present in EGFR protein. Thank you! Any questions? Carrie Iwema iwema@pitt.edu 412-383-6887 Ansuman Chattopadhyay ansuman@pitt.edu 412-648-1297 http://www.hsls.pitt.edu/guides/genetics