Microarray sampling-platform fabrication using bubble-jet technology for a biochip system.
The fabrication of microarrays containing PCR-amplified genomic DNA extracts from mice tumors on a Zetaprobe membrane using a modified thermal ink-jet printer is described. A simple and cost-effective procedure for the fabrication of microarrays containing biological samples using a modified bubble-jet printing system is presented. Because of their mass-produced design, ink-jet printers are a much cheaper alternative to conventional spotting techniques. The usefulness of the biochip microarray platform is illustrated by the detection of human fragile histidine triad (FHIT), a tumor suppressor gene. Subcutaneous carcinomas were induced with MKN/FHIT and MKN/E4 cell lines in immunodeficient mice. Several weeks into their development, the tumors from both groups of mice were removed and subjected to DNA extraction by lysis of tissue samples. The extracted DNA samples were amplified by PCR (30 cycles) using the primers corresponding to nucleotides 2 to 18 of the FHIT sequence. The resulting solution was transferred to the individual reservoirs of a three-color cartridge from a conventional thermal ink-jet printer (HP 694C), and arrays were printed on to a Zetaprobe membrane. After spotting, these membranes were used in a hybridization assay, using fluorescent probes, and detected with a biochip.
Allain
L R
LR
Life Sciences Division, Oak Ridge National Laboratory, TN 37831-6101, USA.
Askari
M
M
Stokes
D L
DL
Vo-Dinh
T
T
D017766:Acid Anhydride Hydrolases; D000818:Animals; D004273:DNA, Neoplasm / Q000032:analysis / Q000302:isolation & purification; D006801:Humans; D051379:Mice; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods*; D009363:Neoplasm Proteins / Q000235:genetics; D009374:Neoplasms, Experimental / Q000175:diagnosis / Q000235:genetics; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000637:transplantation; D000099170:Bis(5'-adenosyl)triphosphatase
D017766:Acid Anhydride Hydrolases; D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D000099170:Bis(5'-adenosyl)triphosphatase