{"product_id":"nxseq-ampfree-low-dna-library-kit-nxgseq-001","title":"NxSeq AmpFREE Low DNA Library Kit","description":"\u003cp\u003eYou’re doing next gen sequencing for a reason; you want big data! To get there, you know you need the best fragment libraries possible to get the most information out of each sample. So why spend your time, resources and expensive sequencing reagents on anything but the best fragment DNA libraries?\u003c\/p\u003e\u003cp\u003eThe NxSeq™ AmpFREE Low DNA Library Kit allows you to build the best fragment DNA libraries possible. We optimised each step of the protocol to ensure peak performance on Illumina sequencers. In addition, these kits require only 75 ng of sheared DNA input, and produce libraries in about 2 hours using a streamlined, easy to follow protocol.\u003c\/p\u003e\u003cp\u003eBetter Data\u003cbr\u003eHigher Efficiency Libraries\u003cbr\u003eMore Reads from Challenging FFPE Samples\u003cbr\u003eExcellent Sequencing Data Examples\u003cbr\u003eMinimal GC Bias\u003cbr\u003eFaster, Easier Protocol\u003cbr\u003eHigher Efficiency Libraries\u003c\/p\u003e\u003cp\u003eMore Sequenceable DNA Fragments per Library = More Data\u003c\/p\u003e\u003cp\u003eFigure 1. 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, Item No.KK4873) and matching amplified library as an internal standard.\u003c\/p\u003e\u003cp\u003eSequencing Impact of Higher Efficiency Library Construction\u003c\/p\u003e\u003cp\u003eBetter Libraries Increase the Number of Reads per Library\u003c\/p\u003e\u003cp\u003eLibrary Kit\tDNA Input\tTotal Number of Sequencing Reads Per Library\u003cbr\u003eStaphylococcus aureus\u003cbr\u003e\tE. coli K12\u003cbr\u003eNxSeq AmpFREE Low DNA Library Kit\t75 ng\t5,649,946\t4,305,882\u003cbr\u003eKapa Hyper Prep Kit\t250 ng\t4,838,726 (-15%)\t1,647,452 (-62%)\u003cbr\u003eIllumina TruSeq DNA PCR-Free Library Prep Kit\t1 µg\t38,768 (-99%)\t1,543,558 (-64%)\u003c\/p\u003e\u003cp\u003eFigure 2a. 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 normalized 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.\u003c\/p\u003e\u003cp\u003eMore Proof with Challenging FFPE Samples\u003c\/p\u003e\u003cp\u003eLibrary Kit\tSample Type\tInput Amount\tTotal Reads\tMapped Reads\u003cbr\u003e(repeat masked)\u003cbr\u003eNxSeq AmpFREE Low DNA Library Kit\tNormal gDNA\t75 ng\t2,163,636\t900,338\u003cbr\u003eFFPE DNA\t75 ng\t1,767,818\t688,074\u003cbr\u003eFFPE DNA\t150 ng\t1,706,714\t656,658\u003cbr\u003eKapa Hyper Prep Kit\tNormal gDNA\t250 ng\t1,567,276 (-28%)\t650,296 (-28%)\u003cbr\u003eFFPE DNA\t250 ng\t1,270,870 (-28%)\t487,872 (-29%)\u003c\/p\u003e\u003cp\u003eFigure 2b. Number of sequencing reads generated from matching normal and FFPE gDNA sample libraries. DNA fragment libraries were prepped using the two indicated kits according to the manufacturer’s recommended input amounts and protocols. Libraries were constructed from normal gDNA (Biochain, Cat. No. D1234142-S02) and DNA extracted from a matching FFPE human kidney tissue (Biochain Cat. No. T2234142-S02) using the Qiagen AllPrep DNA\/RNA FFPE Kit. The gDNA samples were sheared to ~250 bp before starting library construction. Final libraries were quantitated and normalized 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 and mapped reads compared to the corresponding NxSeq AmpFREE Kit results using 75 ng of input DNA.\u003c\/p\u003e\u003cp\u003eHighly Mappable Reads (\u0026gt;92%) from Human, Staphylococcus and Rhodobacter gDNA Sequencing\u003c\/p\u003e\u003cp\u003eSequencing Stat\u003c\/p\u003e\u003cp\u003eHuman\tStaphylococcus\u003c\/p\u003e\u003cp\u003eRhodobacter\u003c\/p\u003e\u003cp\u003eGenome size, GC percentage\u003c\/p\u003e\u003cp\u003e~3 Gbp 45% GC\u003c\/p\u003e\u003cp\u003e2,821,361 33% GC\u003c\/p\u003e\u003cp\u003e4,602,977 69% GC\u003c\/p\u003e\u003cp\u003eRaw reads\u003c\/p\u003e\u003cp\u003e3,131,114\u003c\/p\u003e\u003cp\u003e1,260,836\u003c\/p\u003e\u003cp\u003e3,900,174\u003c\/p\u003e\u003cp\u003eMapped reads\u003c\/p\u003e\u003cp\u003e2,979,237 (95.15%)\u003c\/p\u003e\u003cp\u003e1,174,111 (93.12%)\u003c\/p\u003e\u003cp\u003e3,613,165 (92.64%)\u003c\/p\u003e\u003cp\u003eRead length\u003c\/p\u003e\u003cp\u003e148.9 bp\u003c\/p\u003e\u003cp\u003e148.8 bp\u003c\/p\u003e\u003cp\u003e149.6 bp\u003c\/p\u003e\u003cp\u003eTotal bases\u003c\/p\u003e\u003cp\u003e443,767,447\u003c\/p\u003e\u003cp\u003e174,694,261\u003c\/p\u003e\u003cp\u003e540,403,552\u003c\/p\u003e\u003cp\u003eGenome fraction\u003c\/p\u003e\u003cp\u003e0.11\u003c\/p\u003e\u003cp\u003e0.97\u003c\/p\u003e\u003cp\u003e1.00\u003c\/p\u003e\u003cp\u003eAvg. coverage\u003c\/p\u003e\u003cp\u003e0.15X\u003c\/p\u003e\u003cp\u003e62X\u003c\/p\u003e\u003cp\u003e117X\u003c\/p\u003e\u003cp\u003eFigure 3. 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 normalized 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.\u003c\/p\u003e\u003cp\u003eMinimal Bias Detected\u003c\/p\u003e\u003cp\u003eFigure 4. Sequencing bias measured for three different organisms with varying GC content. DNA fragment libraries were generated from gDNA of three organisms with varying GC content ( Staphylococcus aureus, 24% GC; E. coli K12, 50% GC; and Rhodobacter sphaeroides, 68% GC) according to the manufacturer’s recommended input amounts and protocols. Samples were quantitated using the Bioanalyzer and Qubit fluorometer and normalized to 2 nM final concentrations. Five µL of each library sample was sequenced on a MiSeq using 2 x 150 bp v2 chemistry and analysed. Normalized coverage was calculated as the (average coverage of all windows with X% GC content) divided by the (overall average coverage).\u003c\/p\u003e\u003cp\u003eDon’t spend 25-74% More per Library with Another Kit\u003c\/p\u003e\u003cp\u003eSave Big with the NxSeq AmpFREE Low DNA Library Kit\u003c\/p\u003e\u003cp\u003ePCR-Free Fragment Library Kit\tCat. No.\tSize\u003cbr\u003e(rxns)\t2016 US\u003cbr\u003eList Price\tCost\/Reaction\tPer Rxn Cost Increase\u003cbr\u003evs. NxSeq Kit\u003cbr\u003eNxSeq\u003cbr\u003eAmpFREE Low\u003cbr\u003eDNA Library Kit\t14000-1\t12\t$240\t$20\u003cbr\u003e14000-2\t48\t$921\t$19\t12 rxn\t48 rxn\u003cbr\u003eKapa Hyper Prep Kit\tKK8501\t8\t$264\t$33\t+65%\t+74%\u003cbr\u003eKK8503\t24\t$696\t$29\t+45%\t+53%\u003cbr\u003eKK8505\t96\t$2400\t$25\t+25%\t+32%\u003cbr\u003eFaster Protocol with Significantly Less Hands-on Time\u003c\/p\u003e\u003cp\u003eSave Time and Get Your Samples on the Sequencer Sooner!\u003c\/p\u003e\u003cp\u003eread more\u003c\/p\u003e\n\u003ch2\u003eKey features\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003eLow Input: Requires as little as 75 ng of sheared input DNA allowing use of limiting samples.\u003c\/li\u003e\n\u003cli\u003eHigh Efficiency: Optimised adaptor ligation produces more sequenceable fragments in each library, yielding better coverage \u0026amp; depth from single or multiplexed libraries.\u003c\/li\u003e\n\u003cli\u003ePCR-free: Prevents the introduction of PCR-bias, providing more uniform coverage.\u003c\/li\u003e\n\u003cli\u003eFast: 2 hour, 10 minute protocol saves you time and gets your samples on the sequencer sooner.\u003c\/li\u003e\n\u003cli\u003eAffordable: Best priced and best performing kit available.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003ch2\u003eProduct Details\u003c\/h2\u003e\n\u003cp\u003eThe NxSeq AmpFREE Low DNA Library Kit is compatible with our NxSeq Adaptors or other dT-tailed, Illumina-compatible adaptors.\u003c\/p\u003e\u003cp\u003eEach NxSeq AmpFREE Low DNA Library Kit contains Enzyme Mix (EM), 2X Buffer (2XB), Ligase (LIG) and Elution Buffer (EB). Adaptors must be purchased separately.\u003c\/p\u003e\n\u003ch2\u003eResources\u003c\/h2\u003e\n\u003cp\u003eBrochures and guides\u003c\/p\u003e\u003cp\u003ecase study mcgill nxseq ampfree\u003c\/p\u003e\u003cp\u003ePosters\u003c\/p\u003e\u003cp\u003eNxSeq Fragment Library AmpFREE Low Input PAG 2016\u003c\/p\u003e\n\u003ch2\u003eFAQs\u003c\/h2\u003e\n\u003cp\u003eFAQs\u003c\/p\u003e\n\u003ch2\u003eDocuments\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/sds_NXGSEQ-001_SDS-001-14000-14300-1-14400-1-NxSeq-AmpFREE-Low-DNA-Library-Kit\"\u003eSDS 001 14000 14300 1 14400 1 NxSeq AmpFREE Low DNA Library Kit\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/sds_NXGSEQ-001_SDS-14000-NxSeq-AmpFREE-Low-DNA-Library-Kit\"\u003eSDS 14000 NxSeq AmpFREE Low DNA Library Kit\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/manual_NXGSEQ-001_NxSeq-AmpFREE-Low-DNA-Library-Kit\"\u003eNxSeq AmpFREE Low DNA Library Kit\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/manual_NXGSEQ-001_NxSeq-UltraLow-DNA-Library-Kit-Quick-Protocol\"\u003eNxSeq UltraLow DNA Library Kit Quick Protocol\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/pis_NXGSEQ-001_F103-US-NxSeq-AmpFREE-Kit\"\u003eF103 US NxSeq AmpFREE Kit\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/guide_NXGSEQ-001_case-study-mcgill-nxseq-ampfree\"\u003ecase study mcgill nxseq ampfree\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/poster_NXGSEQ-001_NxSeq-Fragment-Library-AmpFREE-Low-Input-PAG-2016\"\u003eNxSeq Fragment Library AmpFREE Low Input PAG 2016\u003c\/a\u003e\u003c\/li\u003e\n\u003c\/ul\u003e","brand":"Biosearch Technologies","offers":[{"title":"12 Reactions","offer_id":45986428878927,"sku":"14000-1","price":1936.0,"currency_code":"ILS","in_stock":true},{"title":"48 Reactions","offer_id":45986428911695,"sku":"14000-2","price":7357.0,"currency_code":"ILS","in_stock":true},{"title":"fragment","offer_id":45986428944463,"sku":"FRAG","price":710.0,"currency_code":"ILS","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0712\/0748\/5519\/files\/NxSeq-AmpFREE-Low-DNA-Library-Kit_e20534fc-a72c-42f9-947a-df7235099132.jpg?v=1785956067","url":"https:\/\/www.danyel.co.il\/products\/nxseq-ampfree-low-dna-library-kit-nxgseq-001","provider":"danyel biotech","version":"1.0","type":"link"}