41.5 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.
A. |
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 single nucleotide that contains a blocking group on the 3’-OH is added to the array with a cross-linking reagent. 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. The microarray is developed to contain a covalent linker that will dock the DNA oligonucleotide to the array. A new mask is placed on the array and the cycle is repeated until oligonucleotides of the desired length are generated. Unbound nucleotides are washed away from the array. A mask is placed on the array. |
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