Construction and Application of Genomic DNA Libraries

Abstract

Crop failures, pathogenic outbreaks and famine are serious problems facing society. Understanding an organism's genome will help provide the genetic tools needed to solve these complex problems in a shorter time and with less effort. To efficiently study an organism's genome, it can be partitioned into a permanent and stable collection of DNA fragments, called a library. Such libraries provide convenient access to a genome for both laboratory and breeding applications. Genomic libraries can be used as substrates to physically map and sequence entire genomes, clone agriculturally important genes and to investigate gene expression patterns. Further, genomic libraries also provide powerful tools and resources for evaluating germplasm conservation stocks and biological diversity. The ongoing explosion of genetic data and molecular clone resources has opened new scientific possibilities to researchers venturing across all avenues of applied science. In this chapter we will present the details for small and large insert library construction and their applications.

The Korea Brassica Genome Project: A glimpse of the Brassica genome based on comparative genome analysis with Arabidopsis

“…Shotgun sequencing libraries were constructed in pCUGIblu31 for average insert size of 3 kb (Kim et al, 2004;Yang et al, 2004;Yang et al, 2005). BigDye terminators chemistry v3.0 (ABI) was used for the reactions.…”

Toward closing rice telomere gaps: mapping and sequence characterization of rice subtelomere regions

“…Two HaeIII clones, pb083I20 and pb027O22, fortuitously showed the correct HaeIII site (cc) by starting with cc of the telomere motif. The resulting analyses indicate that most of the distal telomere sequences were illegitimately ligated into the EcoRV-digested blunt end vector for the HaeIII library, whereas, it is likely that a fraction of the cohesive ends of the BamHI-digested cloning vector were damaged during preparation of dephophorylated linear vector (Yang et al 2003;Kim et al 2004) (e.g., mechanical breakage or exonuclease contamination of the restriction enzyme or CIP), resulting in blunt ends that would be suitable for telomere cloning. Distal telomere ends are known to contain a single G overhang.…”

Generation of High-Value Genomic Resource in Rice: A “Subgenomic Library” of Low-Light Tolerant Rice Cultivar Swarnaprabha

“…Because genomic libraries represent collections of clones that cover the repertoire of the genomic DNA of an organism, they are useful genetic resources for the physical mapping and cloning of agronomically important genes. Further, genomic libraries are shared as community genomic resources amongst researchers for evaluating germplasm and biological diversity [ 31 ]. In this study, we constructed a subgenomic library from a low-light tolerant cultivar of rice that can be used for the isolation and characterization of multiple light-responsive and intrinsic yield genes, including photoreceptors from rice.…”