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Nucleic Acids Res ; 30(16): e79, 2002 Aug 15.
Article in English | MEDLINE | ID: mdl-12177307

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.


Subject(s)
DNA/metabolism , Oligonucleotide Array Sequence Analysis/instrumentation , Oligonucleotide Array Sequence Analysis/methods , Animals , Candida albicans/genetics , Cell Line , DNA/genetics , DNA/isolation & purification , DNA Primers/genetics , DNA Primers/isolation & purification , DNA Primers/metabolism , Desiccation , Drosophila melanogaster/genetics , Fluorescent Dyes , Gene Library , Glass , Humans , Nucleic Acid Hybridization , Oligonucleotide Array Sequence Analysis/economics , Open Reading Frames/genetics , Polylysine , Polymerase Chain Reaction , Pseudomonas putida/genetics , Quality Control , RNA, Messenger/genetics , RNA, Messenger/isolation & purification , RNA, Messenger/metabolism , Sensitivity and Specificity , Silanes , Time Factors , Trypanosoma brucei brucei/genetics
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