The NEBNext FS DNA Library Prep Kit for MGI is designed for use in MGI sequencing applications. This kit includes optimized components and protocols for the production of high-quality DNA libraries from a wide range (0.1 – 500 ng) of intact DNA inputs, with integrated enzymatic fragmentation, end-repair, and dA-tailing steps.
The NEBNext FS DNA Library Prep Kit is used in conjunction with NEBNext Multiplex Oligos for MGI (Dual Index Primer Pairs Set 1) (NEB #E9725) and libraries are circularized using the NEBNext Circularization Module for MGI (NEB #E9720) before sequencing.
Fast workflow
High library yields
Broad input range: 0.1 – 500 ng
Uniform GC coverage
Lower duplicates and chimeras
Minimized adaptor dimers
Request a sample of this product by visiting the NEBNext Sample Request Page and choosing “Sample not listed” at the bottom of the list; your local NEB Account Manager will reach out to confirm the details of your request.
Product Information
The NEBNext FS DNA Library Prep Kit for MGI meets the need for reliably high-quality DNA library prep from intact samples for MGI sequencing applications.
The kit incorporates trusted NEBNext master mixed reagents and protocols, allowing the use of a wide range of intact DNA inputs (0.1 - 500 ng). The integrated enzymatic fragmentation, end-repair, and dA-tailing step offers fragment-size tunability with a single enzyme mix; the "FS" stands for Fragmentation System.
Note that adaptors, primers, and circularization reagents are provided separately in the NEBNext Multiplex Oligos for MGI (Dual Index Primer Pairs Set 1) (NEB #E9725) and NEBNext Circularization Module for MGI (NEB #E9720).
Get more of what you need with high library yields
Generate high-quality libraries with flexible input amounts from 0.1 ng to 500 ng of intact DNA
Appreciate the convenience of a single adaptor-dilution recommendation
Optimize your time with streamlined workflows, minimized hands-on time, and automation compatibility
Rely on robust performance
FIGURE 1: NEBNext FS DNA Library Prep Kit for MGI workflow
The NEBNext FS DNA Library Prep Kit for MGI has a streamlined workflow with minimal hands-on time across a range of inputs from 0.1 ng to 500 ng.
FIGURE 2: The NEBNext FS DNA Library Prep Kit for MGI enables robust library preparation over a wide input range without adaptor dimers
Libraries were prepared from 0.1–500 ng of human gDNA (NA12878, Coriell Institute for Medical Research) with the indicated PCR cycles. Size selection was performed for a 250 bp average fragment size, except for the 0.1 ng input, where bead cleanup was conducted to avoid excess sample loss and to preserve library complexity. Stock adaptor concentration was used for all input amounts except for 0.1 ng, where a 10-fold dilution of the stock was applied. No traces of adaptor dimer were observed (at ~150 bp) at all input levels (A). Robust library yields were obtained across input amounts. Each bar represents the average of 8–18 replicates, with error bars indicating standard deviation (B). M = Marker
FIGURE 3: The NEBNext FS DNA Library Prep Kit for MGI produces high-quality libraries with excellent sequencing metrics across the input range
Libraries were prepared in duplicate from 0.1–500 ng of NA12878 gDNA (Coriell Institute for Medical Research), sequenced on an MGI® G99 platform with PE100 and downsampled to 3.5 million paired-end reads. Reads were mapped using Bowtie2 (version 2.3.2.2) to the GRCh38 reference, and duplicates were marked using Picard MarkDuplicates (version 1.56.0). Library quality metrics were assessed using Picard Alignment Summary Metrics (version 1.56.0). High mapping rates, low chimera rates, and low adaptor-dimer rates were obtained for all libraries. A slightly higher adaptor-dimer presence (~1%) was observed with the 1 ng input for this specific experiment, and this can be optimized by adaptor dilution. Stock adaptor concentration was used for all input levels except for 0.1 ng, where a 10-fold dilution of the stock was applied.
FIGURE 4: The NEBNext FS DNA Library Prep Kit for MGI produces high-quality libraries with uniform GC coverage across the input range
Libraries were prepared in duplicate from 0.1–500 ng of NA12878 gDNA (Coriell Institute for Medical Research), sequenced on an MGI® G99 platform with PE100 and downsampled to 3.5 million paired-end reads. Reads were mapped using Bowtie2 (version 2.3.2.2) to the GRCh38 reference, and duplicates were marked using Picard MarkDuplicates (version 1.56.0). GC coverage was calculated using Picard's CollectGCBiasMetrics (1.56), which was run on human autosomes only, due to the even copy-number assumption of the tool. Uniform GC coverage was achieved for all input amounts. In the GC coverage plot, the horizontal grey lines indicate the expected normalized coverage of 1.0, and the dots in shades of green represent read numbers at each GC%. The grey area plots below are histograms representing the distribution of GC content in 100 bp windows of the reference genome.
FIGURE 5: The NEBNext FS DNA Library Prep Kit for MGI produces high-quality libraries with consistent insert sizes across a broad input range
Libraries were prepared in duplicate from 0.1–500 ng of NA12878 gDNA (Coriell Institute for Medical Research) with a 250 bp size selection performed for 1–500 ng, or a simple bead cleanup step for 0.1 ng to preserve library complexity. Libraries were pooled and sequenced on an MGI G99 platform with 2x100 base reads and downsampled to 3.5 million paired-end reads. Reads were mapped using Bowtie2 (version 2.3.2.2) to the GRCh38 reference, and insert size distributions were calculated using Picard CollectInsertSizeMetrics (v1.56.0). Consistent insert sizes of 250 bp were achieved for all input amounts with size selection. In contrast, a smaller size distribution was observed for inputs of 0.1 ng where a bead-based clean-up step was used to remove adaptor dimers and excess adaptors, without size selection.
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