5.6 Synthesizing an Oligonucleotide Array
The represents the gene expression patterns of a specific cell type at a given time.
In the DNA microarray methodology, small oligonucleotides are chemically synthesized and are then attached to a very fine grid using a robotic printing device.
In the DNA microarray methodology, oligonucleotides are synthesized directly on the microarray chip using UV-masking and light-directed combinatorial synthesis.
True or False: It is possible to use in-situ synthesized DNA microarray technologies to produce arrays with 65,000 to 100,000 oligonucleotides.
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B. |
True or False: Each nucleotide added to the array needs to be protected so that unwanted extension of the oligonucleotide does not occur.
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B. |
True or False: Once a mask is placed on the array, the protecting groups of the unmasked cells are removed with a chemical lysing reagent.
A. |
B. |
True or False: Arrays containing longer oligonucleotides allow for more accurate and specific identification of gene products.
A. |
B. |
Click on the step and the associated process to identify the series of steps required to construct an in-situ synthesized DNA microarray.
Step 1 Step 2 Step 3 Step 4 Step 5 Step 6 Step 7 | A new mask is placed on the array and the cycle is repeated until oligonucleotides of the desired length are generated. The microarray is developed to contain a covalent linker that will dock the DNA oligonucleotide to the array. A mask is placed on the array. Unbound nucleotides are washed away from the array. The protecting group is removed from the unmasked cells by exposing the array to UV light. The array is coated with a protecting group that covalently binds and blocks the linker. A single nucleotide that contains a blocking group on the 3’-OH is added to the array with a cross-linking reagent. |
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