MICROARRAY MEETING 2002: NEW DEVELOPMENTS IN MUTATION DETECTION AND GENE EXPRESSION
Segrate, MI (Italy), April 12, 2002 

Minerva Biotecnologica 2002 December;14(3-4):247-52

Copyright © 2003 EDIZIONI MINERVA MEDICA

lingua: Inglese

Ligase detection reaction (LDR) and universal array (Zip Code): application to DNA genotyping

De Bellis G. ¹, Castiglioni B. ², Bordoni R. ¹, Mezzelani A. ¹, Rizzi E. ¹, Frosini A. ³, Busti E. ³, Consolandi C. ³, Rossi Bernardi L. ³, Battaglia C. ³

¹ Institute of Biomedical Technologies, National Council of Researches, Milan, Italy; ² Institute of Agricultural Biology and Biotechnology, National Council for Researches, Milan, Italy; ³ Department of Biochemical Sciences and Technologies, CISI, University of Milan, Milan, Italy

The Uni­ver­sal DNA micro­ar­ray ­approach has ­been pro­posed for set­ting up sev­er­al dif­fer­ent DNA anal­y­sis. ­Such an ­approach is ­based on an ­arrayed set of oli­go­nu­cleo­tides (the zip­code sequenc­es) hav­ing com­mon ther­mo­di­nam­ic beha­vi­our but ­being unre­lat­ed to any spe­cif­ic DNA ­sequence ­under ­study. ­This pro­ce­dure, ­that can be com­bined to liga­tion, exten­sion or min­i­se­quenc­ing, ­requires the ­design of a set of ­probes spe­cif­ic for ­each tar­get ­sequence (muta­tions, poly­mor­phism, etc.) append­ed ­with com­ple­men­tary Zip­Code (cZip­Code) sequenc­es. ­These ­probes are react­ed ­against a prop­er tar­get in solu­tion in a mul­ti­plexed for­mat ­using the ­above men­tioned enzy­mat­ic reac­tions. The result­ing prod­ucts, bear­ing ­their own cZip­Code are ­addressed to the loca­tion on the micro­ar­ray ­where the cor­re­spond­ing Zip­Code ­sequence has ­been spot­ted. ­Such an ­array is there­fore “Uni­ver­sal” ­being unre­lat­ed to a spe­cif­ic molec­u­lar anal­y­sis. ­Here we ­present the set up and opti­mi­za­tion of ­this pro­ce­dure and its appli­ca­tion to bac­te­ri­al dis­crim­i­na­tion. ­Four pres­en­ta­tions fol­low dem­on­strat­ing the flex­ibil­ity of the Uni­ver­sal ­array for­mat in detect­ing patho­gens and ­GMOs, mon­i­tor­ing bac­te­ri­al diver­sity and typ­ing ­SNPs.