slide1 n.
Download
Skip this Video
Download Presentation
http://www.biomed.miami.edu/arrays/images/agilent\_array.jpg

Loading in 2 Seconds...

play fullscreen
1 / 15

http://www.biomed.miami.edu/arrays/images/agilent\_array.jpg - PowerPoint PPT Presentation


  • 128 Views
  • Uploaded on

Example of a DNA Array (note green, yellow red colors; also note that only part of the total array is depicted). http://www.biomed.miami.edu/arrays/images/agilent_array.jpg. Methods:.

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about 'http://www.biomed.miami.edu/arrays/images/agilent\_array.jpg' - Samuel


Download Now An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
slide1

Example of a DNA Array(note green, yellow red colors;also note that only part of the totalarray is depicted)

http://www.biomed.miami.edu/arrays/images/agilent_array.jpg

slide2

Methods:

Step 1: Create a DNA array (gene “chip”) by placing single-stranded DNA for each gene to be assayed into a separate “well” on the chip.

slide3

cDNAgene 1

cDNAgene 2

cDNAgene 3

cDNAgene 4

cDNAgene 5

DNA Array: Single-stranded copy DNA foreach gene in a different well.

slide4

Methods:

Step 2: Extract mRNA from biological tissues subjected to an experimental treatment and from the same tissue subjected to a control treatment.

Step 3: Make single-stranded DNA from the mRNA using “color coded” nucleotides.

slide5

Extract mRNA from Control Cells

Extract mRNA from Experimental Cells

Make single-stranded cDNA using green nucleotides

Make single-stranded cDNA using red nucleotides

cDNA = complementary DNA (DNA synthesized from RNA)

slide6

Methods:

Step 4: After making many DNA copies of the RNA, extract an equal amount of cDNA from the controls & experimentals and place it into a separate container.

slide7

Control cDNA

Experimental cDNA

slide8

Methods:

Step 5: Extract a smallamount in a pipette.

slide9

Methods:

Step 6: Insert into first well.

slide10

Methods:

Step 7: Extractmore cDNA and …

… insert intosecond well, etc.

slide11

Methods:

Step 8ff: Continue until all wells arefilled.

slide12

Methods:

Step 9: Allow to hybridize, then wash away all single-stranded DNA.

slide13

Result:

Some wells have no color-coded cDNA (no mRNA in either type of cell)

Some wells have only red (i.e., expressed only in experimental cells)

Some wells have only green (i.e., expressed only in control cells)

Some wells have both red and green in various mixtures (expressedin both experimental and control cells)

slide14

Methods:

Step 10: Scan with a laser set to detect the color & process results on computer.

slide15

Only in

controlcells

Mostly in

controlcells

Mostly in

experimentalcells

Only in

experimentalcells

Same inboth cells

Results:

The colors denote the degree of expression in the experimental versus the control cells.

Gene not expressed in control or in experimental cells

ad