title: Genome physical mapping with large-insert bacterial clones by fingerprint analysis: methodologies, source clone genome coverage, and contig map quality.
abstract: Genome physical mapping with large-insert clones by fingerprint analysis is becoming an active area of genomics research. Here, we report two new capillary electrophoresis-based fingerprinting methods for genome physical mapping and the effects of different fingerprinting methods and source clone genome coverage on quality physical map construction revealed by computer simulations and laboratory experiments. It was shown that the manual sequencing gel-based two-enzyme fingerprinting method consistently generated larger and more accurate contigs, followed by the new capillary electrophoresis-based three-enzyme method, the new capillary electrophoresis-based five-enzyme (SNaPshot) method, the agarose gel-based one-enzyme method, and the automatic sequencing gel-based four-enzyme method, in descending order, when 1% or fewer questionable clones were allowed. Analysis of clones equivalent to 5x, 8x, 10x, and 15x genomes using the fingerprinting methods revealed that as the number of clones increased from 5x to 10x, the contig length rapidly increased for all methods. However, when the number of clones was increased from 10x to 15x coverage, the contig length at best increased at a lower rate or even decreased. The results will provide useful knowledge and strategies for effective construction of quality genome physical maps for advanced genomics research.
authors.affiliation: Department of Soil and Crop Sciences and Institute for Plant Genomics and Biotechnology, 2123 TAMU, Texas A&M University, College Station, TX 77843-2123, USA.
authors.forename: Zhanyou
authors.initials: Z
authors.lastname: Xu
authors.forename: Shuku
authors.initials: S
authors.lastname: Sun
authors.forename: Lina
authors.initials: L
authors.lastname: Covaleda
authors.forename: Kejiao
authors.initials: K
authors.lastname: Ding
authors.forename: Aimin
authors.initials: A
authors.lastname: Zhang
authors.forename: Chengcang
authors.initials: C
authors.lastname: Wu
authors.forename: Chantel
authors.initials: C
authors.lastname: Scheuring
authors.forename: Hong-Bin
authors.initials: HB
authors.lastname: Zhang
mesh_terms: D017360:Arabidopsis / Q000235:genetics*; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003198:Computer Simulation; D020451:Contig Mapping; D004247:DNA / Q000378:metabolism; D016172:DNA Fingerprinting; D015723:Gene Library; D005819:Genetic Markers; D016678:Genome; D006801:Humans; D008957:Models, Genetic; D020161:Physical Chromosome Mapping; D015183:Restriction Mapping; D012984:Software
chemical_list: D005819:Genetic Markers; D004247:DNA