Bioo Scientific Launched the NEXTflex™ PCR-Free DNA Sequencing Kit Reducing Bias in NGS Library Prep

Bioo Scientific announced today that it has launched the NEXTflex™ PCR-Free DNA Sequencing Kit, the first commercial kit that completely eliminates the need for amplification during 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 completely 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.

According to Dr. Toloue, Bioo Scientific’s Director of Genomic Research, the key to eliminating PCR was developing optimized enzymes tailored specifically for each step of library prep. "Typical off-the-shelf enzymes do not require modifications if you're going to end up amplifying your product at the end. The real concern is after sequencing when you realize that your reads have degrees of bias or are contaminated with duplicates and dimers. In the new NEXTflex PCR-Free DNA Sequencing Kit we have made specialized improvements to each enzyme at every step in the library prep protocol. “

The NEXTflex™ PCR-Free DNA Sequencing Kit features “Enhanced Adapter Ligation Technology” which results in library preps with a larger number of unique sequencing 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. 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 makes this the most high-throughput kit available. This kit can be used to prepare genomic DNA libraries for sequencing using Illumina®/Solexa® GAII and HiSeq 2000/1000 platforms.