Introduction to VectorNTI - University of Pittsburgh

advertisement
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
Download