Bioo Scientific has launched the Nextflex PCR-free DNA sequencing kit, which is designed to eliminate the need for amplification during next-generation sequencing (NGS) library prep.
Amplifying AT- or GC-rich genomic regions often leads to sequence-biased nucleotide compositions and poses a serious challenge during analysis.
Using specially designed master-mixed enzymes, the Nextflex PCR-free DNA sequencing kit eliminates the need for amplification, enabling better read mapping and a reduction in duplicate sequences, leading to reduced sequence cost, more representative base identities and better de novo assembly.
In assessing bias, the presence of low-quality and high-GC reads often cannot be aligned well against a reference genome.
High-GC-containing profiles tend to shift toward higher GC content, an indication of poor base representation.
Eliminating PCR removes this content bias for more representative reads.
PCR is also the cause for a significant amount of gene duplicates, which increases sequence cost, adapter dimers and noise in cluster detection.
Dr Toloue, director of genomic research at Bioo Scientific, explained that the key to eliminating PCR was developing optimised enzymes tailored specifically for each step of library prep.
He said: 'Typical off-the-shelf enzymes do not require modifications if you are going to end up amplifying your product at the end.
'The real concern is after sequencing when you realise that your reads have degrees of bias or are contaminated with duplicates and dimers.
'In the Nextflex PCR-free DNA sequencing kit we have made specialised improvement to each enzyme at every step in the library prep protocol,' he added.
The Nextflex PCR-free DNA sequencing kit features Enhanced Adapter Ligation Technology, which results in library preps with a larger number of unique screening reads.
By making improvements to the ligation enzymatic mix, the user will now have the ability to perform ligations with longer adapters and expect to see better binding efficiencies.
Bioo Scientific said the Nextflex PCR-free DNA sequencing kit simplifies workflow by using master-mixed reagents and magnetic bead-based cleanup, reducing pipetting and eliminating time-consuming steps in library preparation.
In addition, the availability of up to 48 unique adapter barcodes is claimed to make this the most high-throughput kit available.
The kit can be used to prepare genomic DNA libraries for sequencing using Illumina/Solexa GAII and Hiseq 2000/1000 platforms.
