A simple and robust method for preparation of cDNA nylon microarrays.
DNA array technology has made remarkable progress in recent years and has become an indispensable tool in molecular biology research. However, preparing high-quality custom-made DNA arrays at a reasonable cost is still an important concern because we cannot abandon the use of DNA array systems designed for specific purposes. To address these problems, we here report the use of rolling circle amplification products of cDNA plasmids dissolved in 80% formamide as DNA probes immobilized on a nylon membrane. First, because formamide is practically non-volatile under ambient conditions and nucleic acids are easily dissolved in it, the use of formamide as a DNA solvent ensures that the DNA concentration of the solution will not change during arraying, which often takes several hours to a day depending on the number of DNA spots and arrays to produce. Secondly, the use of rolling circle amplification technology greatly reduced the labor needed to prepare the spotted DNA. The results in this study demonstrate that the introduction of these two modifications in preparation of nylon DNA array greatly improved its quality.
Yamakawa
Hisashi
H
Department of Human Gene Research, Kazusa DNA Research Institute, 2-6-7 Kazusa-kamatari, Kisarazu, Chiba 292-0818, Japan. yamakawa@kazusa.or.jp
Yokoyama
Satoru
S
Hirano
Takeo
T
Kitamura
Hiroshi
H
Ohara
Osamu
O
D003363:Cost Control; D015342:DNA Probes; D005559:Formamides / Q000737:chemistry; D020869:Gene Expression Profiling; D006801:Humans; D008567:Membranes, Artificial; D009757:Nylons; D020411:Oligonucleotide Array Sequence Analysis / Q000191:economics / Q000379:methods*; D010957:Plasmids; D011786:Quality Control; D013048:Specimen Handling
D015342:DNA Probes; D005559:Formamides; D008567:Membranes, Artificial; D009757:Nylons; C031066:formamide