NEBNext Direct® target enrichment is now available for targeted genotyping!
The NEBNext Direct Genotyping Solution delivers cost-effective, high-throughput, sequence-based target genotyping for a wide variety of applications in plant, animal, and human.
Ideal solution for genotyping hundreds to thousands of markers
Reduce costs and streamline workflow through pre-capture pooling of up to 96 samples
Maximize sequencer efficiency through dual barcode sample indexing plus Unique Molecular Identifier
Unparalleled target coverage uniformity through unique capture-based enrichment
Eliminate marker dropouts with finely tuned bait design
Increase sample throughput using 1-day, automatable workflow
Interested in evaluating the NEBNext Direct Genotyping Solution?
The NEBNext Direct Genotyping Solution combines highly multiplexed, capture-based enrichment with maximum efficiency next-generation sequencing to deliver cost-effective, high-throughput, genotyping for a wide variety of applications. Applicable for marker ranges spanning 100-5,000 markers, pre-capture multiplexing of up to 96 samples combined with dual indexed sequencing allows over 3.8 million genotypes in a single Illumina® sequencing run.
Figure 1: NEBNext Direct Genotyping Solution workflow
Figure 2: Passing filter reads across 96 pooled samples
Passing filter reads across 96 tomato DNA samples that were enriched using a genotyping panel consisting of 2,309 publicly available SolCAP markers and the NEBNext Direct Genotyping Solution. 25 ng of purified tomato DNA was used for each sample. Samples were index-tagged and pooled prior to hybridization and libraries were sequenced on an Illumina® MiSeq® with 20 cycles of Read 1 to sequence the 12 base UMI and 8 base sample index, and 75 cycles of Read 2 to sequence the targets.
Figure 3: NEBNext Direct Genotyping Solution demonstrates similar coverage across 96 pooled samples
Mean SNP coverage of 2,309 SolCAP markers across 96 samples. 25 ng of purified tomato DNA was used for each sample. Samples were index-tagged and pooled prior to hybridization and Libraries were sequenced on an Illumina MiSeq with 20 cycles of Read 1 to sequence the 12 base UMI and 8 base sample index, and 75 cycles of Read 2 to sequence the targets.
Figure 4: Specificity of enrichment of NEBNext Direct Genotyping Solution across 96 pooled samples
The percent of passing filter reads mapping to targeted regions demonstrates high specificity across 96 multiplexed samples using the NEBNext Direct Genotyping Solution. 25 ng of purified tomato DNA was used as input for each sample. Samples were index-tagged and pooled prior to hybridization and Libraries were sequenced on an Illumina MiSeq with 20 cycles of Read 1 to sequence the 12 base UMI and 8 base sample index, and 75 cycles of Read 2 to sequence the targets.
Figure 5: Mean Coverage across 2309 markers within a single sample
Histogram of coverage across each of the 2,309 SolCAP markers demonstrates evenness of enrichment across targets and coverage levels sufficient for genotyping calls. These data represent enrichment of a single tomato sample pooled with 95 others prior to hybridization. 25 ng of purified tomato DNA was used for each sample. Samples were index-tagged and pooled prior to hybridization and libraries were sequenced on an Illumina MiSeq with 20 cycles of Read 1 to sequence the 12 base UMI and 8 base sample index, and 75 cycles of Read 2 to sequence the targets.
Reagents Supplied for E9500
The following reagents are supplied with this product:
Store at (°C)
Concentration
NEBNext Direct® GS Fragmentation Buffer
-20
NEBNext Direct® GS Fragmentation Enzyme Mix
-20
NEBNext Direct® GS Indexed 5′ Adaptor Plate
-20
NEBNext Direct® GS 5' Adaptor Ligation Master Mix
-20
NEBNext Direct® GS Stop Solution
4
NEBNext Direct® GS Sample Purification Beads
RT
Reagents Supplied for E9530
The following reagents are supplied with this product:
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Certificate of Analysis
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]
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