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DNA library preparation for low sample input: NxSeq

The NxSeq® AmpFREE Low DNA library kit enables the preparation of the high-quality, PCR-free fragment DNA libraries from 75 ng of sheared DNA input in just over 2 hours. Utilising a PCR-free protocol and high-efficiency adaptor ligation, these kits produce more sequencable fragments in each library, yielding better coverage and depth from single or multiplexed libraries. Featuring a streamlined, easy-to-follow protocol, these kits are optimised to ensure peak performance on Illumina® sequencers.

Benefits of using the NxSeq AmpFREE Low DNA library kits
• Low input requirement - as little as 75 ng of sheared input DNA
• High efficiency adaptor ligation ensures better coverage & depth
• Quick and easy protocol - library prep in just over 2 hours
• Compatible with a wide range of samples including genomic DNA, FFPE and limited samples
• Optimised for Illumina® sequencers
• PCR-free protocol prevents PCR-bias for a more uniform coverage
• Cost efficient option for low input samples

The NxSeq AmpFREE Low DNA library kit produces higher efficiency libraries than other commercially available kits

The NxSeq AmpFREE Low DNA library kit produces higher efficiency libraries than other commercially available kits

Percentage of library DNA with correctly ligated adaptors measured by qPCR. Duplicate libraries were prepped per kit/organism (Human, Staphylococcus aureus, Rhodobacter sphaeroides (1 library only), and E. coli) according to the manufacturer’s recommended input amounts and protocols. Adaptor ligation efficiency was measured by qPCR using the KAPA™ Library Quantification Kit (Complete ROX Low, cat #KK4873) and matching amplified library as an internal standard.

The NxSeq AmpFREE Low DNA library kit offers highly mappable reads (>92%) from Human, Staphylococcus and Rhodobacter gDNA sequencing

Sequencing stat Human Staphylococcus Rhodobacter
Genome size, GC percentage ~ 3 Gbp, 45% GC 2,821,631, 33% GC 4,602,977, 69% GC
Raw reads 3,131,114 1,260,836 3,900,174
Mapped reads 2,979,237 (95.15%) 1,174,111 (93.12%) 3,613,165 (92.64%)
Read length 148.9 bp 148.8 bp 149.6 bp
Total bases 443,767,447 174,694,261 540,403,552
Genome fraction 0.11 0.97 1.00
Average coverage 0.15X 62X 117X


Representative gDNA sequencing stats from three different organisms. Genomic DNA fragment libraries were generated using the NxSeq AmpFREE Low DNA Library Kit using 75 ng of sheared gDNA input from three organisms (human, Staphylococcus aureus, and Rhodobacter sphaeroides). The final libraries were quantitated and normalised to 2 nM final concentrations using the Bioanalyzer® and Qubit™ fluorometer, and 5 µL of each library was run on a MiSeq® using 2 x 150 bp chemistry and analysed.

Material available for download
NxSeq AmpFREE Low DNA Fragment Library kit manual

Illumina, MiSeq and TruSeq are registered trademarks of Illumina, Inc.
KAPA is a trademark of Roche Molecular Systems, Inc. 
Bioanalyzer is a registered trademark of Agilent Technologies, Inc. 
Qubit is a registered trademark of Thermo Fisher Scientific.

Products

Note: product availability depends on country - see product detail page.

Details Cat number & supplier Size Price
NxSeq™ AmpFREE Low DNA Library Kit 14000-1 · Lucigen
14000-1
Lucigen
12 reactions £338.00
12 reactions
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NxSeq™ AmpFREE Low DNA Library Kit 14000-2 · Lucigen
14000-2
Lucigen
48 reactions £1290.00
48 reactions
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NxSeq Adaptors, Box 1 14300-1 · Lucigen
14300-1
Lucigen
12 x 4 reactions £245.00
12 x 4 reactions
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NxSeq™ Adaptors, Box 2 14400-1 · Lucigen
14400-1
Lucigen
12 x 4 reactions £245.00
12 x 4 reactions
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