NxSeq® 40 kb Mate-Pair Cloning Kit

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CAT#	PRODUCT	SIZE	STORAGE	소비자가	주문
42028-2	NxSeq™ 40kb Mate-Pair Cloning Kit	10 rxns	-20º & -80º C	-	로그인
42028-3	NxSeq™ 40kb Mate-Pair Cloning Kit	20 rxns	-20º & -80º C	-	로그인

NxSeq® 40 kb Mate-Pair Cloning Kit

True long-span, mate-pair libraries for Next Generation Sequencing

Complete your genome with 40 kb, long-span, mate-pair NGS libraries
Skip the adapters: pNGS vector contains built-in Illumina and Roche 454 primers sites
Ideal for de novo, whole genome sequencing projects
Transcription free vector: highly stable clones

Introducing a new paradigm in long-span, mate-pair Next Gen libraries:
the NxSeq 40kb Mate-Pair Cloning Kit.

The pNGS FOS vector (fig. 1) represents the combination of Lucigen's patented transcription free vectors with a first-of-its-kind feature: built-in Roche 454 and Illumina primer binding sites – no adapter ligation necessary! Using a 40 kb paired-end library, you can now build the critical scaffold that enables a truly complete genome assembly. Get exceptionally stable clones due to our transcription-free vector, reduce time and uncertainty by skipping adapter ligation, and get more sequence coverage with less bias than ever before.

Figure 1.

Unique Workflow:
Let Lucigen make the process of generating long-span, mate-pair libraries easier than ever before.

Shear and size select for 40 kb fragments, ligate to the pNGS FOS vector and perform lambda packaging.
Cut with a 4bp restriction enzyme (RsaI, CviQI, BfaI, or FspBI)
Reconstitute the restriction site to create a known mate-pair junction.
Amplify with either Roche 454 or Illumina primers and proceed to sequencing (fig. 2).

Figure 2.

High Efficiency 40 kb Paired-end Sequencing for Next Generation Platforms

To demonstrate the efficiency of cloning and sequencing using the pNGS FOS vector, large-scale, long-span, mate-pair sequencing of a human cell line (GM15510, Coriell) using Illumina technology resulted in 64% of filtered reads accurately mapping to the genome (Table 1). This represents many-fold higher efficiency than existing systems and will allow the accurate assembly of genomes for the first time using next gen sequencing platforms.

Table 1. Summary of Illumina long-span, mate-pair sequencing of a human cell line
Stage	No. of Reads	%
Pipeline	8,683,854
Filtered reads	6,506,126
Reads mapped	5,173,778	79.2
Reads with pair-sequences	4,685,996	90.6
Long-span, mate pairs*	3,018,576	64.4
*Mapped on same chromosome, ~40 kb apart, uniquely mapped.

Genotypes

Replicator FOS Strain: F¯ mcrA Δ(mrr-hsdRMS-mcrBC) endA1 recA1 Φ80dlacZΔM15 ΔlacX74 araΔ139 Δara,leu)7697 galU galK rpsL nupG (attL araC-PBAD-trfA250 bla attR) λ-.

MA113_NxSeq-40kb-Mate-Pair-Manual.pdf (1,888,381kb)

MSDS.pdf (118,131kb)

NGS-FOS-long-span-mate-paire.pdf (708,858kb)

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관련제품: NxSeq® AmpFREE Low DNA Library Kits and Adaptors (Illumina-compatible)