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Lecture 20

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0 views8 pages

Lecture 20

Uploaded by

Nagarakshita V K
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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DNA Microarrays

What is a DNA Microarray?


 Also known as DNA Chip
 Allows simultaneous measurement of the
level of transcription for every gene in a
genome (gene expression)
 Transcription?
 Process of copying of DNA into messenger RNA
(mRNA)
 Environment dependant!
 Microarray detects mRNA, or rather the more
stable cDNA
2
What is a DNA Microarray?
(cont.)

Cheung et al. 1999

Patrick Schmid 3
How do we manufacture a
microarray?
 Start with individual genes, e.g. the ~6,200
genes of the yeast genome
 Amplify all of them using polymerase chain
reaction (PCR)
 “Spot” them on a medium, e.g. an ordinary
glass microscope slide
 Each spot is about 100 µm in diameter
 Spotting is done by a robot
 Complex and potentially expensive task

Patrick Schmid 4
How do we manufacture a
microarray?

Cheung et al. 1999

Patrick Schmid 5
Example

 Remember the flash animation?


 Yeast
 Grow in aerobic and anaerobic environment
 Different genes will be activated in order to
adapt to each environment
 Extract mRNA
 Convert mRNA into colored cDNA
(fluorescently labeled)

Patrick Schmid 6
Example (cont.)

 Mix cDNA together


 Hybridize cDNA with array
 Each cDNA sequence hybridizes specifically
with the corresponding gene sequence in the
array
 Wash unhybridized cDNA off
 Read array with laser
 Analyze images

Patrick Schmid 7
Overview of Example

Brown & Botstein, 1999

Patrick Schmid 8

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