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Probes/Targets DNA Arrays...Probes: are the tethered nucleic acids with known sequence, –the DNA on the chip,...Target: is the free nucleic acid sample.

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Presentation on theme: "Probes/Targets DNA Arrays...Probes: are the tethered nucleic acids with known sequence, –the DNA on the chip,...Target: is the free nucleic acid sample."— Presentation transcript:

1 Probes/Targets DNA Arrays...Probes: are the tethered nucleic acids with known sequence, –the DNA on the chip,...Target: is the free nucleic acid sample whose identity/abundance is being detected, –the labeled nucleic acid that is washed over the chip.

2 Figure 1d

3

4 Figure 1a, DNA ChipFigure 1b, DNA Microarray Probe? Target? Oligos Yeast Transcripts Under Two Conditions. Probe?cDNAs

5 Signal to Noise, Data? Establish noise levels, Detect single colors alone to determine expression yes/no questions. Compare ratios of red to green to establish relative expression levels.

6 Hybridization Detection …fluorescent images are read by an optical scanner, and intensities are compared using algorithms to differentiate artifacts.

7 Figure 2

8 Fig. 3

9 Figure 4

10 Figure 5

11 Figure 6

12 Probe? Target? Adult retina cDNA library wt mice? cDNA libraries from Crx +/+ and Crx -/- 10 day postnatal mice.

13 Control #1...one cDNA library,...split, 1/2 Cy3, 1/2 Cy5,... + 1/1 correlation between signals.

14 Control #2...one brain polyA-mRNA,...two reverse transcriptases,...one labeled with Cy3, one with Cy5,... + 1/1 correlation between signals.

15 Replicates #1 and #2... targets Crx +/+ and Crx -/- 10 day postnatal mice.,...two reverse transcriptases,...one labeled with Cy3, one with Cy5,... + 1/1 correlation between signals.

16 Replicate #1... targets Crx +/+ and Crx -/- 10 day postnatal mice,... some points fall off of the + 1/1 regression....high on the Cy5 side.

17 Table #1

18 Figure #3 Northern Blot, –RNA on substrate, –probed with “hot DNA”.

19 Table #2 What does this mean? Does it really?

20 Figure #4 Probe? Target (a.)? Adult retina cDNA library wt mice? adult mouse retina prenatal mouse retina Target (b.)? adult retina adult brain

21 Figure #5 (a) spot 501, (b) spot 140, (c) calcium channel (Crx +/+ ), (d) (c) calcium channel (Crx -/- ).

22 Figure #6


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