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This vid briefly explains Phosphoramidite.

i. Phosphoramidite chemistry is commonly used in the synthesis of nucleic acids, particularly in the production of synthetic oligonucleotides, which are short sequences of nucleotides (DNA or RNA) used in various molecular biology applications, such as PCR, sequencing, and gene editing.

In phosphoramidite chemistry, a phosphoramidite reagent is used as a key building block for oligonucleotide synthesis. This reagent typically consists of a nucleoside base (adenine, guanine, cytosine, or thymine/uracil) attached to a phosphoramidite group via a linker molecule. The phosphoramidite group allows for controlled addition of nucleotides during oligonucleotide synthesis.

During the synthesis process, the phosphoramidite reagent is coupled to a growing oligonucleotide chain anchored to a solid support. This coupling reaction involves the activation of the phosphoramidite group, typically with a tetrazole catalyst, followed by reaction with a hydroxyl group on the growing oligonucleotide chain. This process results in the formation of a phosphite triester linkage between the incoming nucleotide and the growing chain.

After each coupling step, unreacted phosphoramidite reagents and byproducts are typically removed, and the cycle is repeated to add subsequent nucleotides until the desired oligonucleotide sequence is obtained.

Overall, phosphoramidite chemistry provides a highly efficient and versatile method for synthesizing custom-designed oligonucleotides with precise control over sequence composition and length, making it a fundamental technique in molecular biology and biotechnology.