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9 Cards in this Set
- Front
- Back
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What molecule is measured in "gene expression"?
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mRNA (transcript)
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What are some of the methods used to determine the expression of "a handful of genes"?
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1. Northern Blot Hybridization
2. Slot Blot Hybridization 3. RT-PCR |
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What are some of the methods used to measure the expression of a large number (thousands) of genes?
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1. High Throughput Northern Blot Analysis
2. Differential Display Analysis 3. Subtractive Hybridization |
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What is the underlying principle in microarray technology?
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Hybridization (base Pairing)
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In MPSS, how is sequencing used to determine gene expression?
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Massively Parallel Signature Sequencing
1. Unique 32 nucleotide tags are attached to cDNAs and the cDNA with tags are amplified by PCR. 2. Many copies of anti-tag which is complementary to the 32-nt tags are attached to beads. Each bead has a unique anti-tag. 3. cDNA-tags from step 1 are incubated with the beads. Complementary tags and anti-tags attach to each other by complementary base-pairing. 4. Beads are immobilized in a flow cell. Each individual bead is coated with about 100,000 copies of the amplified signature from one mRNA transcript. The number of beads corresponds to the prevalence of the mRNA molecules. 5. Counting of the beads yields the frequency of a particular mRNA in the sample. 6. From each cell 20-bp signature sequences are generated. |
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What is meant by gene expression?
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Gene expression is the process of creating a gene product. This gene product creation process starts with DNA to create mRNA that in turn is used to produce protein or other gene product.
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What are the different ways by which gene expression can be measured?
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Depending on the amount mRNA present a number of processes exist for measuring gene expression including RT-PCR, SAGE, MicroArrays, and Northern Blot.
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How is microarray technology different from SAGE technology?
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SAGE uses rDNA to clone randomly linked short sequences whereas Microarray technology depends on hybridization. SAGE does not depend on having foreknowledge of the which sequences are being looked for nor does it require extensive expression of the gene.
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What are some of the advantages of MPSS?
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Massively Parallel Signature Sequencing does not depend upon prior knowledge of the genome or of the expressed genes, so it can be used for gene discovery as well as expression analysis. (As compared to Microarrays)
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