Will Nanopore Sequencing Make it Obsolete?
Genome sequencers have been tweaking polymerase chain reaction (PCR) amplification to avoid introducing artifacts into sequencing libraries, ranging from modifications in chemistries to the introduction of novel sequencing technologies that could obviate the need for PCR altogether.
PCR-related problems have included uneven amplification, causing overrepresentation of some sequence species and nucleotide misincorporation. In particular, sequencing genomes or genomic regions with extremely biased base composition remains a challenge to the currently available next-generation sequencing (NGS) platforms. These include, for example, the genomes of important pathogenic organisms like Plasmodium falciparum with a high adenine–thymine (AT) content and Mycobacterium tuberculosis with high guanine–cytosine (GC) content.
These genomes have proven difficult for sequencers because the standard library preparation procedures that employ PCR amplification cause uneven read coverage across these regions, leading to problems in genome assembly and variation analyses.
Read more: PCR-Free Novel Genome Sequencing Technology
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| Disruptive sequencing based on nanopore technology has the potential to change completely the way DNA sequencing is done. |
Source: beawolf/Fotolia
