Manufacturing DNA microarrays from unpurified PCR products.

نویسندگان

  • Frank Diehl
  • Boris Beckmann
  • Nadine Kellner
  • Nicole C Hauser
  • Susanne Diehl
  • Jörg D Hoheisel
چکیده

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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عنوان ژورنال:
  • Nucleic acids research

دوره 30 16  شماره 

صفحات  -

تاریخ انتشار 2002