Mate-Pair NGS Libraries for Genome Assembly and Gap Closure
• Complete your genome with 40 kb, long-span, mate-pair NGS libraries
• pNGS vector contains built-in Illumina, Roche 454, or Ian Torrent primers sites
• Ideal for de novo, whole genome sequencing projects
• Transcription free vector gives highly stable clones
Large insert mate-pair reads have a major impact on the overall success of de novo genome assembly and the discovery of inherited and acquired structural variants. Molecular tools that bridge the gap between massively parallel short read sequencing technologies (35-1,500 bases) and the need for large scaffolds (>40,000 bases) to accurately assemble complex repeat-rich genomes are required. The NxSeq 40 kb Mate-Pair Cloning Kit has been developed to facilitate genome assembly and gap closure.
Using this kit approximately 40 kb paired-end sequences can be obtained by either Illumina, 454 or Ion Torrent sequencing at an overall efficiency of 70%, which is significantly better than existing long-span, mate-pair systems.
The NxSeq 40 kb Mate-Pair Cloning system utilizes a unique fosmid vector design. Fosmid vectors are important tools for positional cloning, physical mapping, and genomic sequencing. In fosmid cloning, DNA fragments of ~35-45 kb are cloned into a fosmid vector and packaged into phage particles in vitro by bacteriophage Lambda packaging extract. The phage particles are then used to insert the fosmid DNA into bacteria, which typically maintain it as a single-copy plasmid until replication is induced.
Most fosmid vectors yield small amounts of DNA and are prone to instability due to transcriptional interference. Our pNGS FOS vector minimizes transcription both into and out of the insert DNA, reducing the cloning bias found with conventional vectors. Most importantly, the pNGS vector contains primer binding sites for Next Generation Sequencing (NGS) from Illumina, Roche 454 or Ion Torrent platforms.
Nature Methods Application Note (Online Version)
Poster Presentation about 40 kb Mate-Pair Kit
NxSeq DNA Sample Prep Kits
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