NEBNext® Ultra™ II FS DNA PCR-free Library Prep Kit for Illumina®
Don’t be biased! The NEBNext Ultra II FS DNA PCR-free Library Prep Kit for Illumina offers an amplification-free workflow for DNA-seq based on the streamlined and reliable Ultra II FS workflow. Starting with as little as 50 ng of DNA, get the high-quality, high-yield libraries that you need without PCR bias.
Includes NEBNext FS (Fragmentation System) enzymatic fragmentation reagents
Perform fragmentation, end repair and dA-tailing with a single enzyme mix
Save time with an optimized PCR-free workflow, reduced hands-on time and automation compatibility
Generate high quality libraries without an amplification step using inputs from 50 ng to 500 ng intact DNA
The NEBNext Ultra II FS DNA PCR-free Library Prep Kit contains the enzymes and buffers required to convert a broad range of intact DNA input amounts into libraries for next-generation sequencing on the Illumina platform, all without an amplification step. The fast, user-friendly workflow includes reliable enzymatic DNA fragmentation, requires minimal hands-on time, resulting in excellent library yields and GC coverage without requiring PCR.
Advantages
Enjoy flexible input amounts from 50 ng to 500 ng of intact DNA
Experience reliable fragmentation with a single protocol, regardless of DNA input amount or GC content
Perform fragmentation, end repair and dA-tailing with a single enzyme mix
Get more of what you need with reliably high library yields
Prepare PCR-free libraries with a broad range of sample types
Save time with streamlined workflows, reduced hands-on time, and automation compatibility
Use with the NEBNext Multiplex Oligos for Illumina (Unique Dual Index UMI Adaptors DNA Set 1) (NEB #E7395) or other Illumina-compatible index adaptors with a 3’ single T overhang
Figure 1: NEBNext Ultra II FS DNA PCR-free Library Preparation Kit generates libraries with higher yields
A. PCR-free libraries were prepared with human cell line NA19240 genomic DNA (Corielle Institute) using NEBNext Ultra II FS DNA PCR-free and Illumina PCR-free library prep kits and size selected for 350 bp long inserts.
B. Libraries of 150-200 bp inserts were prepared using the enzyme-based fragmentation kits NEBNext Ultra II FS DNA PCR-free and Roche Sequencing Kapa HyperPlus PCR-free library prep kits without size selection. NEBNext Unique Dual Index UMI Adaptors DNA Set 1 and Kapa Dual-Indexed Adaptors were used for the NEBNext and Kapa kits, respectively, following manufacturers' recommendations.
Figure 2: NEBNext Ultra II FS DNA PCR-free Library Prep provides uniform genome coverage with human DNA
NEBNext Ultra II FS DNA PCR-free libraries have more even genome coverage than Illumina DNA PCR-free. Sequencing libraries were prepared according to the manufacturer's recommendations from 250 ng of NA19240 cell line DNA by 2 different users and sequenced on a single NovaSeq® 6000 S4 v1.5 flow cell (PE 2x150 bp). Reads were adaptor trimmed (fastp 0.20.0), aligned to the Telomere-to-Telomere chm13 references (draft 1, bwa-mem 0.7.17), and duplicate marked (samblaster 0.1.24). 2.8B reads were randomly sampled from each bam (sambamba view -s 0.6.8). Coverage of primary, pass-filter assignments was assigned using mosdepth (v0.3.1, -F 772) and plotted for autosomes or the mitochondrial genome (inset). Expected coverage (num_reads*read length/genome size) is indicated. There is significant variation between users for Illumina PCR-free libraries; however, the FS DNA PCR-free libraries are very consistent between users.
This product is related to the following categories:
Use this tool to find citations related to NEBNext products. Search by product, keyword or instrument name – use the filters to select specific product areas.
Quality, Safety & Legal
Quality Control Assays
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications & Change Notifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
Based on your Freezer Program type, you are trying to add a product to your cart that is either not allowed or not allowed with the existing contents of your cart. Please review and update your order accordingly. If you have any questions, please contact Customer Support at freezers@neb.com or 1-800-632-5227 x 8.
You have been idle for more than 20 minutes, for your security you have been logged out. Please sign back in to continue your session.
Institution Changed
Your profile has been mapped to an Institution, please sign back for your profile updates to be completed.
Sign in to your NEB account
To save your cart and view previous orders, sign in to your NEB account. Adding products to your cart without being signed in will result in a loss of your cart when you do sign in or leave the site.
Want to try this product? Request a sample
Need a custom/large volume order? Contact us
