NxSeq® AmpFREE Low DNA Library Kit

This PCR-free, low input fragment library preparation kit has an optimised protocol to ensure peak performance on Illumina® sequencers. The high efficiency, PCR-free NxSeq® AmpFREE Low DNA Library Kit enables the use of low input amounts without the need for PCR amplification prior to sequencing.

Benefits of using the NxSeq® AmpFREE Low DNA Library Kit
• High efficiency library construction
• Around two hour protocol
• Simple protocol
• Over 75 ng sheared DNA required
• Minimal bias - PCR-free
 

% 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.

Better libraries increase the number of reads per library

Library kit	DNA input	Total number of sequencing reads per library (Staphylococcus aureus)	Total number of sequencing reads per library (E. coli K12)
NxSeq® AmpFREE Low DNA Library Kit	75 ng	5,649,946	4,305,882
Kapa Hyper Prep Kit	250 ng	4,838,726 (-15%)	1,647,452 (-62%)
Illumina TruSeq DNA PCR-Free Library Prep Kit	1 µg	38,768 (-99%)	1,543,558 (-64%)

Number of sequencing reads generated per library after multiplexing and running on a MiSeq Instrument. DNA fragment libraries were prepped in parallel for each kit/organism according to the manufacturer’s recommended input amounts and protocols. Libraries were quantitated and normalised to 2 nM using the Bioanalyzer (size) and Qubit Fluorometer (amount). Equimolar amounts of each library were multiplexed and sequenced with a single MiSeq run using 2 ×150 bp chemistry. The number of sequencing reads obtained are shown as well as the percent reduction (%) in total reads compared to the appropriate NxSeq AmpFREE Kit results.

Faster protocol with significantly less hands-on time

 

The NxSeq® AmpFREE Low DNA Library Kit and Adaptors are only compatible with Illumina sequencers. Each NxSeq® AmpFREE Low DNA Library Kit contains Enzyme Mix (EM), 2X Buffer (2XB), Ligase (LIG) and Elution Buffer (EB). Adaptors must be purchased separately.
Each box of NxSeq® Adaptors contains 12 different indexed Illumina-compatible adaptors with enough of each adapter for four library reactions. Box One contains adaptors 1-12 and Box Two contains adapters 13-24.

NxSeq® AmpFREE Low DNA Library Kit index sequence

Applications
• De novo genome sequencing
• Resequencing
• Single nucleotide polymorphism (SNP/mutation) detection

Material available for download
NxSeq® AmpFREE Low DNA Library Kit Manual
NxSeq® AmpFREE Low DNA Library Kit Product Flyer
NxSeq® AmpFREE Low DNA Library Kit Poster

Illumina is a registered trademark and MiSeq a trademark of Illumina, Inc

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