Canadian Forest Service Publications
Constructing conifer genomic libraries: a basic guide. 1989. DeVerno, L.L.; Byrne, J.R.; Pitel, J.A.; Cheliak, W.M. Forestry Canada, Petawawa National Forestry Institute, Chalk River, Ontario. Information Report PI-X-88E. 26 p.
Available from: Atlantic Forestry Centre
Catalog ID: 6876
Series: Information report (Petawawa)
Procedures are given for constructing genomic DNA bacteriophage libraries from conifer tissue. Although black spruce Picea mariana (Mill.) B.S.P. tissue was used, the protocols can be applied to any plant species. Included are techniques for high molecular weight genomic DNA isolation from mature needles, seedlings, and cell suspension cultures. In addition, purification of size-fractionated genomic DNA fragments, vector DNA isolation, and preparation of vector arms are outlined. Details are provided for production of recombinant clones, in vitro packaging, amplification, and evaluation of library quality. Appendices contain methods for purification of sized fragments, media and buffer formulas, and the number of recombinants required to represent genomes of various species.