DNA microarrays: Difference between revisions
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DNA microarrays are used in [[ | [[DNA|DNA]] microarrays are used in [[Functional genomics|functional genomics]] to determine the differences in [[Gene expression|gene expression]] levels between a sample and control [[cell|cell]] <ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. The sample cell can be: from a different tissue, at a different stage of development, at a different stage of the [[Cell cycle|cell cycle]], or be under different conditions (for example, exposure to a [[Toxin|toxin]]) <ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. The DNA microarray consists of a flat surface to which [[Oligonucleotides|oligonucleotides]] are bound <ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. These oligonucleotides are complementary to specific [[CDNA|cDNA]] sequences <ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. The [[MRNA|mRNA]] [[Molecules|molecules]] within the sample and the control are converted into labelled cDNA molecules with the use of [[Reverse transcriptase|reverse transcriptase]] and fluorescently-labelled [[Nucleotide|nucleotides]]<ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. | ||
For example, the cDNA of the sample can have a red fluorescence label whereas the cDNA of the control can have a green fluorescence label <ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. The DNA microarray is exposed to the cDNA mixture and unbound cDNA is washed away <ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. The resultant DNA microarray consists of spots of colour that is imaged using a [[Confocal fluorescence scanner|confocal fluorescence scanner]] <ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. The colour of the spot is indicative of the differences in [[gene|gene]] expression between the sample and control <ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. Following the colour scheme above, a red spot indicates that the sample is overexpressing that particular [[Gene|gene]] compared to the control; a green spot indicates that the sample is underexpressing that particular gene compared to the control; and a yellow spot indicates that there is equal gene expression in the sample and control <ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. However, the range of colours is not as discrete as suggested here, it is more of a spectrum covering intermediate differences in gene expression<ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. DNA microarrays are not so useful in determining gene function but can ascertain which genes may have the same regulatory mechanisms<ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. | |||
For example, the cDNA of the sample can have a red fluorescence label whereas the cDNA of the control can have a green fluorescence label<ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. The DNA microarray is exposed to the cDNA mixture and unbound cDNA is washed away<ref name="Daniel L. Hartl">Daniel L. Hartl, Elizabeth W. Jones (2009) Genetics Analysis of Genes and Genomes 7th Edition USA, Jones and Bartlett Publishers.</ref>. The resultant DNA microarray consists of spots of colour that is imaged using a [[ | |||
=== References === | === References === | ||
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Revision as of 20:53, 7 December 2011
DNA microarrays are used in functional genomics to determine the differences in gene expression levels between a sample and control cell [1]. The sample cell can be: from a different tissue, at a different stage of development, at a different stage of the cell cycle, or be under different conditions (for example, exposure to a toxin) [1]. The DNA microarray consists of a flat surface to which oligonucleotides are bound [1]. These oligonucleotides are complementary to specific cDNA sequences [1]. The mRNA molecules within the sample and the control are converted into labelled cDNA molecules with the use of reverse transcriptase and fluorescently-labelled nucleotides[1].
For example, the cDNA of the sample can have a red fluorescence label whereas the cDNA of the control can have a green fluorescence label [1]. The DNA microarray is exposed to the cDNA mixture and unbound cDNA is washed away [1]. The resultant DNA microarray consists of spots of colour that is imaged using a confocal fluorescence scanner [1]. The colour of the spot is indicative of the differences in gene expression between the sample and control [1]. Following the colour scheme above, a red spot indicates that the sample is overexpressing that particular gene compared to the control; a green spot indicates that the sample is underexpressing that particular gene compared to the control; and a yellow spot indicates that there is equal gene expression in the sample and control [1]. However, the range of colours is not as discrete as suggested here, it is more of a spectrum covering intermediate differences in gene expression[1]. DNA microarrays are not so useful in determining gene function but can ascertain which genes may have the same regulatory mechanisms[1].
References