UNIT 5.20 Fosmid Libraries for Genomic Structural Variation Detection
Published Online: 1 JUL 2007
Copyright © 2007 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Human Genetics
How to Cite
Donahue, W. F. and Ebling, H. M. 2007. Fosmid Libraries for Genomic Structural Variation Detection. Current Protocols in Human Genetics. 54:5.20:5.20.1–5.20.18.
- Published Online: 1 JUL 2007
- Published Print: JUL 2007
Fosmid libraries have demonstrated their utility for a number of applications. These include filling gaps between BACs and small insert libraries in sequence assemblies, performing hybridization/screening studies to isolate functional elements within the genome (Vergin et al., 1998), and detecting insertions, deletions, and rearrangements in structural variation studies (Tuzun et al., 2005). This unit covers the basic methodologies for the construction of fosmid libraries with tight insert sizes suitable for these applications. Basic Protocol 1 covers the shearing, size selection, and recovery of DNA from a pulsed-field gel. Basic Protocol 2 covers the cloning of insert DNA into the fosmid vector, packaging of DNA into infective phage particles, and the infection/transformation of bacteria. A commentary section is provided, which outlines many of the critical parameters involved in fosmid library construction, along with some additional background information and a section discussing anticipated results. Curr. Protoc. Hum. Genet. 54:5.20.1-5.20.18. © 2007 by John Wiley & Sons, Inc.
- fosmid cloning;
- fosmid library;
- fosmid protocol;
- structural variation detection