Laboratoryof Molecular Medicine, Instutute of Medical Science, the University of Tokyo
Manufacture protocol
36,864 transcripts including ESTs were chosen as target cDNAs, and their sequences were retrieved from the UniGene database (National Center for Biotechnology Information; build# 131). A mixture of pooled mRNA from the liver, spleen, thyroid, placenta, skeletal muscle, small intestine, brain, heart, fetal lung, fetal liver, fetal kidney, and fetal brain (Clontech) were used for target cDNA preparation. Each RNA was reverse transcribed using oligo(dT) primer and Superscript II reverse transcriptase (Invitrogen). We amplified cDNA segments of 200-1100-bp long without repetitive or polyadenylated sequences by PCR using the specific primer sets to each gene or EST. The 36,864 PCR products were purified and spotted in duplicate on type 7 or Alice glass slides (Amersham Pharmacia Biotech) using a Microarray Spotter Generation III (Amersham) or Lucidea Microarray Spotter (Amersham), separated on eight or four slides. Each slide contained 52 housekeeping genes, to normalize the signal intensities of the different fluorescent dyes. Spotted DNAs were cross-linked with slides by SPECTRO LINKER (SPECTRONIC Corp.).
Description
Human genome-wide cDNA microarray with 36,864 elements spotted (sets1-8)
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