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Author: Malloff, C
Author: Lam, W
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Methods Article

Two-dimensional DNA displays for comparisons of bacterial genomes

Chad Malloff1, Edie Dullaghan2, Alice Li3, Richard Stokes4, Rachel Fernandez5* and Wan Lam1

1 Departments of Pathology and Laboratory Medicine. British Columbia Cancer Research Center. Vancouver, BC. Canada.
2 Department of Pediatrics and the Division of Infectious and Immunological Diseases. British Columbia Children's Hospital. Vancouver, BC. Canada.
3 Departments of Pathology and Laboratory Medicine. The Division of Infectious and Immunological Diseases. British Columbia Children's Hospital. Vancouver, BC. Canada.
4 Departments of Pathology and Laboratory Medicine, Pediatrics, Microbiology and Immunology. University of British Columbia. The Division of Infectious and Immunological Diseases. British Columbia Children's Hospital. Vancouver, BC. Canada.
5 Microbiology and Immunology. University of British Columbia. Vancouver, BC. Canada.

* To whom correspondence should be addressed: Rachel Fernandez, Department of Microbiology & Immunology University of British Columbia. Vancouver, BC. Canada. Phone: (604) 822-6824. Fax: (604) 822-6041. Email: rachelf@interchange.ubc.ca

Biol. Proced. Online 2003;5:143-152. doi:10.1251/bpo56
Submitted: March 16, 2003; Accepted: April 01, 2003; Published: June 15, 2003.

Indexing terms: Genomics; DNA fingerprinting.


Figure 2 Enlarged

Fig. 2:

Schematic outlining Bacterial Comparative Genomic Hybridization. (a) Samples to be compared are digested with the same frequent cutting enzyme(s) and combined. (b) The combined sample is then size separated on a first dimension polyacrylamide gel. (c) The first dimension gel lane is then cut from the gel and transferred to the top of a denaturing gradient polyacrylamide gel. (d) Sample fragments are then resolved in the second dimension based on melting characteristics. (e) The gel is then washed and the DNA electroblotted to a nylon membrane. (f,g) The nylon membrane is then alternatively hybridized with radiolabelled genomic DNA probe (asterisk) of generated from the comparison samples.

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