DNA microarrays (or “gene chips”) contain thousands of oligonucleotides anchored

Question

DNA microarrays (or “gene chips”) contain thousands of oligonucleotides anchored to a solid surface in defined locations. Gene chips enable the rapid visualization and quantification of changes in gene expression across complex samples (including entire organisms). The DNA strands on these chips are synthesized using iterative phosphoramidite couplings as discussed in lecture. Each “spot” on a gene chip contains several copies of a unique DNA sequence; the collection of these “spots” can represent a large fraction of the genome for an organism.
a. A portion of the gene encoding for SNCA is shown below. SNCA encodes for alpha-synuclein, an abundant brain protein involved in neurotransmission. SNCA overexpression has been linked to Parkinson’s disease in some patients. Describe a synthetic method to produce the oligonucleotide (on solid support) that could be selectively attached to a solid surface (e.g., a gene chip) in a second step. (5'-GGTAGGTGAATCTTTC-3')

b. Assuming that you had the DNA strand but did not know its identity. Design an experiment that will allow you to sequence the DNA strand.

Explanation / Answer

1. Complimentary strand of DNA is synthesized by PCR method and the produced strand is immbilised due to charge difference between DNA and immobilising surface on the chip which hybrised with the gene and inactivates to cure the disease.

2. For Seqencing DNA first attempt was made by Sanger and used dideoxynucleotides lebelled and complimentry binding causes sequencing. Other method like PCR methods FISH also may used for DNA sequencing.

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