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Example sentences for: fragments
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The cDNA array has a long history of development [ 3 ] stemming from immunodiagnostic work in the 1980s; however, it has been most widely developed in recent years by Stanford University (California) researchers depositing cDNA tags onto glass slides, or chips, with precise robotic printers [ 4 ] . Labeled cDNA fragments are then hybridized to the tags on the chip, scanned, and differences in mRNA between samples identified and visualized using a variation of the red/green matrix originally introduced by Eisen and colleagues [ 5 ] . The light-generated oligonucleotide array, developed by Affymetrix, Inc. (Santa Clara, CA), involves synthesizing short 25-mer oligonucleotide probes directly onto a glass slide using photolithographic masks [ 6 7 ] . Sample processing includes the production of labeled cRNA, hybridization to a microarray, and quantification of the obtained signal after laser scanning.
DNA fragments coding for peptides containing 40 random amino acids were generated by a PCR approach using synthetic oligonucleotides as previously described [ 7 26 ] . Peptides are expressed on the capsid protein pIII of the phage at low copy number (1-2 peptides/phage).
In this microarray, 7075 immobilized cDNA fragments (4107 from known genes and 2968 ESTs) were immobilized onto a glass slide.
(This number is slightly less than the number of retained fragments (153) because, whereas most of the retained fragments were flanked by lost regions, a few directly bordered other retained fragments.)
Cycle sequencing extension products were created in a final volume 20 μl reaction using 8 μl H 2 O, 4 ul forward or reverse primer at 20 ng/μl, 4 μl template DNA (amplicon) at 4 ng/μl for fragments up to 200 bp, 5 ng/μl for fragments between 200 and 400 bp and 6 ng/μl for fragments greater than 400 bp, 2 μl Big Dye Terminator Ready Reaction mix (PE-ABI, Foster City, Ca.)
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