Preparation of DNA and protein micro arrays on glass slides coated with an agarose film.
A thin layered agarose film on microscope slides provides a versatile support for the preparation of arrayed molecular libraries. An activation step leading to the formation of aldehyde groups in the agarose creates reactive sites that allow covalent immobilization of molecules containing amino groups. Arrays of oligonucleotides and PCR products were prepared by tip printing. After hybridization with complementary fluorescence labeled nucleic acid probes strong fluorescence signals of sequence-specific binding to the immobilized probes were detected. The intensity of the fluorescence signals was proportional to the relative amount of immobilized oligonucleotides and to the concentration of the fluorescence labeled probe. We also used the agarose film-coated slides for the preparation of protein arrays. In combination with specific fluorescence labeled antibodies these protein arrays can be used for fluorescence linked immune assays. With this approach different protein tests can be performed in parallel in a single reaction with minimal amounts of the binding reagents.
Afanassiev
V
V
Hans-Knöll-Institut für Naturstoff-Forschung, Beutenbergstrasse 11, D-07745 Jena, Germany.
Hanemann
V
V
Wölfl
S
S
D002352:Carrier Proteins / Q000378:metabolism; D015723:Gene Library; D005898:Glass; D006801:Humans; D008904:Miniaturization; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000378:metabolism; D019151:Peptide Library; D012685:Sepharose; D051025:bcl-Associated Death Protein
C494819:BAD protein, human; D002352:Carrier Proteins; D009841:Oligonucleotides; D019151:Peptide Library; D051025:bcl-Associated Death Protein; D012685:Sepharose