19a microarrays

859 views
505 views

Published on

Published in: Technology
0 Comments
1 Like
Statistics
Notes
  • Be the first to comment

No Downloads
Views
Total views
859
On SlideShare
0
From Embeds
0
Number of Embeds
51
Actions
Shares
0
Downloads
0
Comments
0
Likes
1
Embeds 0
No embeds

No notes for slide

19a microarrays

  1. 1. Microarrays Microarrays are used to examined thousands of mRNA strands all at the same time. Two samples: 1. Control (normal / wild type) 2. Treatment (e.g. chemical) / Abnormal (e.g. cancer)
  2. 2. Microarrays  Up to 100 000 pieces of known DNA sequence are placed onto the slide  The position of each DNA sequence is known and kept in a database
  3. 3. Microarrays
  4. 4. Steps to Microarray Analysis 1. mRNA is isolated from tissue 2. cDNA is made using mRNA as template (using reverse transcriptase) cDNA will also be labeled with a fluorescent protein   e.g. control = green; drug treatment = red 3. fluorescent cDNA is placed onto the microarray 4. laser scanner to locate fluorescence of each sample
  5. 5. cDNA Microarray Assay estrogen in sesame oil (vehicle) sesame oil harvest tissue extract RNA Incorporate Cy3 label Incorporate Cy5 label Mix and hybridize Scan Data Collection Data analysis
  6. 6. Microarray Animation
  7. 7. Scanned Microarray
  8. 8. Scanned Microarray 1. Green  more mRNA produced in control sample 2. Red  more mRNA produced in treated sample 3. Yellow  both control and treated sample produce the same amount of mRNA
  9. 9. Microarray Information What do you do with all these colours? Focus on the green or red dots  these represent mRNA that are different between the control and the test sample If you know which gene is being changed, you may understand which metabolic pathway is being affected.

×