Literature DB >> 12177307

Manufacturing DNA microarrays from unpurified PCR products.

Frank Diehl1, Boris Beckmann, Nadine Kellner, Nicole C Hauser, Susanne Diehl, Jörg D Hoheisel.   

Abstract

For the production of DNA microarrays from PCR products, purification of the the DNA fragments prior to spotting is a major expense in cost and time. Also, a considerable amount of material is lost during this process and contamination might occur. Here, a protocol is presented that permits the manufacture of microarrays from unpurified PCR products on aminated surfaces such as glass slides coated with the widely used poly(L-lysine) or aminosilane. The presence of primer molecules in the PCR sample does not increase the non-specific signal upon hybridisation. Overall, signal intensity on arrays made of unpurified PCR products is 94% of the intensity obtained with the respective purified molecules. This slight loss in signal, however, is offset by a reduced variation in the amount of DNA present at the individual spot positions across an array, apart from the considerable savings in time and cost. In addition, a larger number of arrays can be made from one batch of amplification products.

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Year:  2002        PMID: 12177307      PMCID: PMC134252          DOI: 10.1093/nar/gnf078

Source DB:  PubMed          Journal:  Nucleic Acids Res        ISSN: 0305-1048            Impact factor:   16.971


  12 in total

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  8 in total

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