Chapter_14_-_Post-genomic_analysis_v2

Chapter_14_-_Post-genomic_analysis_v2 - Post-genomic...

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Post-genomic Analysis
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14.1  Analysing transcription; transcriptomes 14.1.1 Differential screening Compare two different mRNA populations, using the different populations as cDNA probes Hybridize different probes to duplicate library filters Positive signals with one probe but not the other are candidate clones of differentially expressed genes Drawbacks to differential screening method: Differentially expressed genes (high and low abundance) are not necessarily reflected in a differential intensity of signals on the filter Cannot pick up expressed genes of low abundance Figure 14.1 Differential screening of a gene library
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14.1  Analysing transcription; transcriptomes  14.1.2 Subtractive hybridization Subtractive hybridzation is the removal of common sequences from the two different mRNA populations the driver (control) cDNA removed from the tester cDNA Driver cDNA labeled with biotin Driver cDNA forms biotin linkage to streptavidin beads Tester cDNA forms hybrids upon reannealing to Driver cDNA (which is provided in excess of Test cDNA) Hybrid tester-driver cDNA bound to stretavidin beads are removed, leaving an enriched population of unique Tester cDNA Figure 14.2 Subtractive hybridization
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14.1  Analysing transcription; transcriptomes  14.1.2 Representational difference analysis (RDA) Representational difference analysis is a PCR based method that requires less starting mRNA than traditional subtractive
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This note was uploaded on 07/10/2010 for the course BIOL 208 taught by Professor Chuong during the Spring '09 term at Waterloo.

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Chapter_14_-_Post-genomic_analysis_v2 - Post-genomic...

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