Choose your NEBNext Workflow
Select a workflow
to see the respective formats and how it builds up on the Ultra II DNA workflow
End Repair/ dA-Tailing
Adaptor Ligation
Clean Up/ Size Selection
PCR Enrichment
Clean Up
component of #E7103
Module #M0544
Module #E7595
Module #E7546
NEBNext Ultra II DNA Library Prep Kit for Illumina (#E7645)
NEBNext Enzymatic Methyl-seq Workflow
NEBNext Ultra II FS DNA Library Prep Workflow
NEBNext Single Cell/Low Input RNA Workflow
NEBNext Ultra II Directional RNA or Non-directional RNA Workflow
NEBNext Ultra II DNA Library Prep Kit for Illumina The NEBNext Ultra II reagents and kits for Illumina platforms compose the core of your NGS library preparation: a perfect combination of reagents, optimized formulations and simplified workflows. With incredibly short “hands-on” time you create libraries of highest quality and yields even when starting from extremely low input quantities.
Hands on time: <15 minutes Total time: 2:30 – 3:00 hrs
NEBNext Ultra II DNA Workflow
NEBNext ARTIC SARS-CoV-2 FS Library Prep Workflow
For more information, please visit www.neb.com/E7645
Input amounts: 500 pg – 1 µg total DNA
Enzymatic DNA Fragmentation
&
Module #E7810
NEBNext Ultra II FS DNA Library Prep Kit (#E7805)
NEBNext Ultra II FS DNA Library Prep Kit for Illumina The Ultra II FS DNA kit includes a unique 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.
Hands on time: <15 minutes Total time: ~ 2:30 hrs
Input amounts: 100 pg – 0.5 μg unsheared DNA
For more information, please visit www.neb.com/e7805
RNA fragmentation and priming
Poly(A) enrichment
rRNA depletion
OR
First strand cDNA synthesis
Second strand cDNA synthesis
NEBNext Ultra II directional or non-directional RNA Library Prep Kits (#E7760 and #E7770)
Module #E6150
Module #E7490 or various depletion kits
Module #E7771
Module #E7550 or #E6111
NEBNext Ultra II Directional and Non-Directional RNA Library Prep Kit for Illumina The NEBNext Ultra II RNA kits are available as directional and non-directional workflows. After polyA selection or depletion of unwanted RNA as well as cDNA synthesis and fragmentation the reagents are following the NEBNext II Ultra II DNA protocol to generate high quality libraries with high yields and even coverage of the transcriptome. The NEBNext Ultra II Directional RNA kit is providing also strand-specific information by using the highly regarded dUTP method for superior data quality.
Hands on time: 15 – 30 minutes Total time: ~ 5:30 – 6:45 hrs
Input amounts: 5 ng – 1 μg total RNA (rRNA depletion) 10 ng – 1μg total RNA (poly(A) mRNA)
For more information on the directional RNA Workflow, please visit www.neb.com/e7760
For more information on the non-directional RNA Workflow, please visit www.neb.com/e7770
Genomic DNA Shearing by Sonication
Oxidation of 5mC and 5hmC
Deamination of C to U
PCR Amplification
not included
Module #E7125
Module #M0597
NEBNext Enzymatic Methyl-seq Kit (#E7120)
NEBNext Enzymatic Methyl-Seq Kit (EM-seq) Whole Genome Bisulfite Sequencing (WGBS) has long been the gold standard for methylome analysis, but the chemical bisulfite reaction damages and degrades DNA, resulting in fragmentation and loss. Additionally, bisulfite libraries demonstrate significant GC bias and are enriched for methylated regions. To overcome these limitations, we developed an enzyme-based approach, NEBNext Enzymatic Methyl-seq (EM-seq), an alternative method for identification of 5mC and 5hmC.
Hands on time: 56 minutes Total time: 6 hrs – 7 hrs
Input amounts: 10 ng – 200 ng
For more information, please visit www.neb.com/e7120
Sample Purification Beads not included
Single Cell Lysis
Reverse Transcription
Template Switching
cDNA Amplification
Enzymatic Fragmentation End Repair/ dA-Tailing
Module #E5530
Module #E6421
NEBNext Single Cell/Low Input RNA Kit (#E6420)
NEBNext Single Cell/Low Input RNA Library Prep Kit for Illumina How low can you go? This unique workflow meets the demand for a highly sensitive, yet robust method that consistently generates high-quality, full-length transcript sequencing data from a single cell or ultra-low input RNA. Optimized cDNA synthesis and amplification steps incorporate template switching, as well as utilize a unique protocol and suite of reagents. Even low-abundance transcripts are represented in the high yields of cDNA obtained. Subsequent library construction incorporates the Ultra II FS enzymatic DNA fragmentation/end repair/dA-tailing mix in a simple and efficient workflow.
Hands on time: <30 minutes Total time: 6 hrs – 7 hrs
Input amounts: 2 pg – 200 ng total RNA
For more information, please visit www.neb.com/e6420
NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina) The NEBNext ARTIC kit meets the increasing need for reliable, accurate and fast methods for sequencing SARS-CoV-2 using Illumina sequencing. The kit uses the ARTIC multiplexed amplicon-based whole-viral-genome sequencing approach and were developed in collaboration with the ARTIC network. The kit includes balanced V3 primer pools for improved uniformity of genome coverage and are effective with 10 – 10,000 SARS-CoV-2 genome copies. By converting the viral Genome into cDNA followed by amplification the kit is building on the NEBNext FS DNA kit for library preparation.
NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (#E7658)
cDNA Synthesis
Module #E7626
Enzymatic Fragmentation
Hands on time: ~ 30 minutes Total time: ~ 8 hrs
Input amounts: 10 – 10.000 viral copies
For more information, please visit www.neb.com/e7658
