While both methodologies delve into DNA analysis at the single-base level, their approaches differ significantly.
Sanger sequencing relies on the pioneering dyetermination technique introduced by Frederic Sanger. This method generates fragments of the target sequence, each initiated from a primer but terminated at varying lengths. Each fragment terminates with a fluorescent label, distinguished by different colors corresponding to the A, C, G, and T nucleotides. These fragments are discerned following separation by capillary electrophoresis, hence also known as capillary electrophoresis sequencing (CES).
On the other hand, NGS encompasses a broad spectrum of techniques for sequencing DNA libraries. These libraries comprise extensive collections of short DNA fragments, sourced from fragmented genomic DNA or RNA, amplicons, or even mixed genome fragments from diverse species (metagenomes). In this high-throughput methodology, all sequences within a library are sequenced collectively, with subsequent bioinformatics analysis used to reconstruct the original sequence(s) comprehensively.
In essence, Sanger sequencing offers a rapid, dependable, and cost-effective approach when examining a limited number of short genomic regions. In contrast, next-generation sequencing (NGS) excels in efficiently exploring extensive genomic regions, such as whole genomes or exomes, leveraging massively parallel sequencing technologies.
Sanger sequencing demonstrates unparalleled accuracy in analyzing sequences ranging from hundreds to thousands of bases. Its precision makes it invaluable for identifying single nucleotide polymorphisms (SNPs) at specific loci, as well as for species identification through the analysis of molecules like 16S rRNA.
Moreover, the stream lined workflow of Sanger sequencing renders it suitable for applications where prompt results are imperative. For instance, at Macrogen Europe, we offer Sanger sequencing with turnaround times as swift as 24 hours.
NGS emerges as the preferred technology for investigating extensive regions of the genome, transcriptome, or epigenome. Its capability for massively parallel sequencing enables the efficient exploration of diverse genomic entities, ranging from targeted gene panels to entire genomes orexomes.
NGS also shines in metagenomics, facilitating the comprehensive study of all species present in a mixed sample. This is particularly beneficial in microbiome analysis, where NGS can identify species using 16S amplicon sequencing or delve into whole genomes with meta-shotgun sequencing.
At Macrogen Europe, we offer an array of Sanger sequencing and NGS services, including massively parallel sequencing. If you are uncertain about the most suitable technique for your requirements, reach out to us now for a complimentary consultation and discover the optimal solution tailored to your needs.