Resource Summary Report  New Search Previous Search Results

URL: https://dx.doi.org/10.17504/protocols.io.p3adqie

Authors: Elena Hilario, Marcus Davy, Roy Storey, Richard Newcomb, Ross N Crowhurst, Roger P Hellens

Summary: A genome is assembled by connecting large scaffolds via a collection of thousands of mate-paired ends of various sizes (3-20 kbp). Terminal ends of long DNA fragments can be obtained by sequencing large insert clones by Sanger sequencing, a labour-intensive and expensive approach. Here we describe a high-throughput protocol to tag 11,520 BAC plasmids randomly with barcoded oligonucleotides at the end of a random hexamer by PCR.This method assisted the kiwifruit (Actinidia chinensis) genome assembly in a similar way as mate paired end datasets.  Although not fully sequenced, each randomly tagged BAC clone provides a collection of short sequence tags mapped to a genome across one or multiple scaffolds under three different scenarios: validate a single scaffold, join two scaffolds, or group several scaffolds by proximity only. Our method provides a simple way of cataloging BAC libraries to assist genome assembly projects.

Affiliations: Plant and Food Research, Plant and Food Research, Plant and Food Research, Plant and Food Research, Plant and Food Research, Queensland University of Technology

External URL: https://bmcgenomics.biomedcentral.com/articles/10.1186/s12864-018-4656-3

Version: 1

Publication Date: 2018

Expand All Collapse