The Illumina-solexa sequencing protocol for bacterial genomes

Abstract

Based on reversible dye-terminators technology, the Illumina-solexa sequencing platform enables rapid sequencing-by-synthesis (SBS) of large DNA stretches spanning entire genomes, with the latest instruments capable of producing hundreds of gigabases of data in a single sequencing run. Illumina’s NGS instruments powerfully combine the fl exibility of single reads with short- and long-insert paired-end reads, and enable a wide range of DNA sequencing applications. Here, we describe the paired-end library preparation with an average insert size of 470 bp, 2 kbp, and 6 kbp, together with the DNA cluster generation and sequencing procedure of E. coli O104:H4 genome on Illumina Hiseq 2000 platform.