NEBNext Ultra II kits are designed for construction of high-quality libraries using a broad range of input amounts, from pg to μg of DNA. The workflows are fast, streamlined and automatable, with flexible product formats to enable customization.
NEBNext Ultra II FS DNA Library Prep – a novel enzymatic fragmentation system
We have built upon our NEBNext Ultra II DNA library prep workflow to create a fragmentation system, the NEBNext Ultra II FS DNA Library Prep Kit. The Ultra II FS kit includes a new DNA fragmentation reagent, which is also combined with end repair and dA-tailing reagents, enabling these steps to be performed in the same tube, with no clean-ups or sample loss. The same fragmentation protocol is used for any input amount (100 pg–500 ng), or GC content.
You'll be thrilled to pieces with the result – a reliable, flexible, high-quality library prep that is fast and scalable.
Highlights of the NEBNext Ultra II FS Kit
- Perform fragmentation, end repair and dA-tailing with a single enzyme mix
- Experience reliable fragmentation with a single protocol, regardless of DNA input amount or GC content
- Prepare high quality libraries from a wide range of input amounts: 100 pg–500 ng
- Generate high yields with increased reaction efficiencies and minimized sample loss
- Use with DNA in standard buffers (TE, Tris-HCl) and water
- Save time with a streamlined workflow: ~ 2.5 hours, with < 15 minutes hands-on time
- Vary incubation time to generate a wide range of insert sizes
- Available with optional SPRIselect® beads for size selection/clean-up
See what others are saying about Ultra II
“In our core, we process large number of samples for genomics applications. Even with the high-throughput Covaris® model, the sample shearing is still a bottle neck for us. The NEBNext DNA DNA kit is such a wonderful product. First of all, it increased our throughput dramatically for DNA sample shearing. The kit combines the shearing, end-repair, dA-tailing, and linker-ligation in a couple simple steps without the need of purification in between. The whole process has been reduced from two days to a few hours. Second, we love the low input option. Because the kit eliminates the need for purification for each of the intermediate steps, it reduces a lot of sample loss during the purification steps, which enables us to use much less starting materials and saves tons on purification beads. Third, we are amazed by the randomness of the enzymatic shearing. We did not observe any significant difference from mechanical shearing. We have used this kit successfully with many large projects including high-throughput genomic screening assays for single cell derived clones.
- Scientist, Cancer research institute
This video walks you through DNA Library Preparation using the NEBNext Ultra II DNA Library Prep Kit. The video also includes tips for optimization as well as safe stopping points.
This video walks you through the size selection and cleanup steps using the NEBNext Ultra II DNA
In this webinar, Laurence Etwiller and Jennifer Ong discuss their recent publications, and how library preparation affects sequencing accuracy and variant calling. View full Q & A summary.
Optimizing DNA inputs for NGS library prep is an important step, if you want to ensure a high-quality library. Start by following these tips for DNA quantification and assessing purity.
This video will help you decide if you need to do a size selection or a simple clean-up for the NEBNext Ultra DNA or RNA.
A workflow animation for an enzyme-based method for detection of 5mC and 5hmC, an alternative to bisulfite sequencing.
In this ESHG 2020 workshop, hear about methylation analysis of cfDNA samples, sample identification and tracking using a high-throughput SNP genotyping method and the importance of sample prep in large-scale sequencing pipelines.
Improving Enzymatic DNA Fragmentation for Next Generation Sequencing Library Construction
Among the available options for DNA fragmentation, enzymatic fragmentation is demonstrably gentler on the sample, yielding less damage at any scale. The new NEBNext® Ultra™ II FS DNA Library Prep Kit for Illumina® includes a single step for enzymatic fragmentation, end repair, and dA tailing.
The Quantitation Question: How does accurate library quantitation influence sequencing?
The determination of the number of sequencing-ready molecules present after library preparation is an important step in the next generation sequencing (NGS) workflow and has a strong influence on the success of both a sequencing run and a sequencing-based experiment.
- Troubleshooting Guide for NEBNext® Ultra™ II FS DNA Library Prep Kit
- A Robust, Streamlined, Enzyme-based DNA Library Preparation Method Amenable to a Wide Range of DNA Inputs (2019)
- NEBNext® Ultra™II FS DNA: A Robust Enzyme-based DNA Library Preparation Method Compatible with Plant and Animal Samples (2019)
- Uncovering the Cannabis sativa Methylome Through Enzymatic Methyl-seq (2019)
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