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Biotechniques

Biotechniques. DNA Chip. Microarrays are small thin wafers of glass / silicon Contain 100s – 1000s of known DNA probes Can identify sequences of unknown target DNA. DNA Chip Applications. determine code of unsequenced DNA ( eg genome analysis)

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Biotechniques

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  1. Biotechniques

  2. DNA Chip Microarrays are small thin wafers of glass / silicon Contain 100s – 1000s of known DNA probes Can identify sequences of unknown target DNA

  3. DNA Chip Applications • determine code of unsequenced DNA (eg genome analysis) • identify alleles in unknown sample (eg detect genetic markers for diseases such as cystic fibrosis ) • map genetic differences (eg genes may have single base difference – can tell people apart)

  4. Single-stranded DNA fragment of known sequence Radioactive or fluorescent dye tag. DNA probes G T G T G T Fluorescent dye tagfluorescent bands when gel is exposed to UV light source. or Radioactive tag dark band when the gel is exposed to photographic film.

  5. Probe tags complementary DNA sequence on target Target DNA strand Probes bind to a complementary sequence on target

  6. Microarrays

  7. DNA Chips DNA probes fixed to a small solid support such as a glass slide or a nylon filter. Each spot on the microarray has thousands to millions of copies of a different DNA probe.

  8. How DNA Chips Work • The tagged sample cDNA will bind with any complementary probe. Such binding indicates that the gene represented by the chip DNA was expressed, or active, in the sample. • After hybridization, the color of the spot indicates the relative amount of mRNA in the samples. The microarray is scanned and a computer quantifies the amount of gene activity in the sample and generates a color-coded read-out.

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