dna microarray n.
Download
Skip this Video
Loading SlideShow in 5 Seconds..
DNA Microarray: PowerPoint Presentation
Download Presentation
DNA Microarray:

Loading in 2 Seconds...

play fullscreen
1 / 13

DNA Microarray: - PowerPoint PPT Presentation


  • 103 Views
  • Uploaded on

DNA Microarray:. A Recombinant DNA Method. Basic Steps to Microarray:. Obtain cells with genes that are needed for analysis. Isolate the mRNA using extraction buffer. Remove the buffer with RNA. The mRNA will be 5’-3’ with repeating AAA..end. Convert each mRNA into colored cDNA.

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 'DNA Microarray:' - melina


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
dna microarray

DNA Microarray:

A Recombinant DNA Method

basic steps to microarray
Basic Steps to Microarray:
  • Obtain cells with genes that are needed for analysis.
  • Isolate the mRNA using extraction buffer.
    • Remove the buffer with RNA.
    • The mRNA will be 5’-3’ with repeating AAA..end.
  • Convert each mRNA into colored cDNA.
    • Targets labeled with fluorophores.
basic steps to microarray1
Basic Steps to Microarray:
  • With mRNA degraded reverse cDNA created.
  • Mix colored samples of cDNA.
microarray or dna chip
Microarray or DNA Chip:
  • Every spot on the chip represents a different coding sequence from different genes.
  • Each spot on the chip is made of a DNA probe that can pair with the cDNA that was created.
basic steps to microarray2
Basic Steps to Microarray:
  • Incubation with the mixed cDNA and the chip DNA will yield some pairing.
  • Wash off unbound cDNA to see what has bound to the microarray.
visualizing bound cdna
Visualizing Bound cDNA:
  • The slide with the microarray chip is placed inside a dark box where it is scanned with a high resolution laser that detects the bound fluorescent labels.
    • The information and images are then sent to the computer for analysis.
analyzing the data
Analyzing the Data:
  • Creates a ratio image.
  • Green images signal expression in one condition.
  • Red images signal expression in one condition.
  • Yellow images signal expression in both conditions.
capacities
Capacities:
  • Not limited to human genetic material.
    • Can be used for all species.
  • Can display thousands of different genes.
    • Pending chip size.
      • 100’s nucleotides (<1 cm^2)
      • 100,000’s nucleotides (about 10 cm^2)
  • Allows the study of multiple genes at once.
types of microarray
Types of Microarray:
  • Pre-synthesized nucleic acids.
    • Bought cDNA.
  • Oligonucleotide synthesis in situ.
    • Creating cDNA in question.
microarray application
Microarray Application:
  • Identifying gene’s and gene mutations of different types of diseases such as cancer.
  • Identifying the expression level, or quantity of genes (mutated or not).
definitions
Definitions:
  • Target - the nucleic acid (cDNA) sample who’s identity and quantity are being measured.
  • Fluorophore – usually green and red labels attached to the target to enable visualizing expression.
  • Microarray works as reverse hybridization method converting from mRNA to cDNA 3’-5’ with TTTT…end.
  • Probe – an attached nucleic acid with a known sequence (the DNA chip).
resources
Resources:
  • Strachan, Tom; Read, Andrew. Human Molecular Genetics 3. pp 175-178. New York. Garland Publishing. 2004.
  • Campbell,A. Malcom. Microarray Animation. Davidson College. 2001. www.bio.davidson.edu/courses/genomics/chip/chip.html
  • http://cmgm.stanford.edu
  • http://www.gene-chips.com/