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Manufacturing DNA microarrays from unpurified PCR products

: Diehl, F.; Beckmann, B.; Kellner, N.; Hauser, N.C.; Diehl, S.; Hoheisel, J.D.

Postprint (PDF; )

Nucleic Acids Research 30 (2002), No.16, pp.E79-E79
ISSN: 0305-1048
ISSN: 0301-5610
ISSN: 1362-4962
Journal Article, Electronic Publication
Fraunhofer IGB ()

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.