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Positive Controls Printed
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On all Mouse,Rat & Human subarrays:
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Operon positive 70 mer control, actin, S19 ribosomal protein or 28S rRNA, & genomic DNA
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On all M.tuberculosis:
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subarrays:Operon positive 70mer control, Sig Factor A, 16s rRNA, & genomic DNA
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Negative Controls Printed
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Operon negative 70 mer controls
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SSC Printing: salt spots w/o DNA
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Blanks: no salt or DNA
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Hybridization & Normalization Controls:
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3 B.subtilis genes printed & cRNAs synthesized- for Reverse transcription, cDNA synthesis and hybridization
controls
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Post Printing: Verification
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Within each array, each subarray will have a common pattern of positive, negative, and salt controls for rapid visual orientation.
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Each array design has a unique pattern of controls: mouse 1, mouse 2, M.tuberculosis, Mouse 3, Human 3 and Rat 1.1.
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Hybridization of random flurescent 9mer to view spot morphology and verify that all features printed.
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Hybridize anti-sense fluorescent 70 mer targets, complimentary to specific mouse 70 mer probes, to prove accurate subarray.
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printing and verify selected probe sequences.
Figures:
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