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  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

language: English

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


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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.

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