Development of a DNA Library Synthesis Protocol for Illumina-Compatible Sequencing of Human Genomic DNA

Abstract

DNA sequencing is a technique used to determine the nucleotide sequence of DNA. There has been a vast advancement in sequencing over the past years, such as employing faster sequencing methods. One of these methods is Next Generation Sequencing, where millions of small DNA fragments are sequenced simultaneously. While several equipment platforms exist with different sequencing methods, one of the most popular systems is that of Illumina, based in San Diego, California (http://www.illumina.com). Illumina sequencing is cost efficient per base-pair of DNA, and sequences the genome faster than previously used methods i.e Sanger’s method. This is a result of the extremely high throughput of the number of bases read over time. A library containing sequenceable DNA is made in order for the DNA to be sequenced via Illumina sequencing. DNA sequencing is widely used to study the cause of genetic disorders like Familial Exudative Vitreoretinopathy (FEVR) and Norrie Disease. For our experiment, we made two duplicate libraries using protocols provided by manufacturers. Lastly, three quality control tests were done during this experiment in order to confirm that sequenceable DNA was synthesized.