신제품 NEBNext UltraExpress™ Library Prep Kit는 권장 입력 범위 내의 샘플에 하나의 어댑터 농도와 PCR 사이클링 조건으로 시간이 단축되어 효율적인 라이브러리 준비에 최적화된 제품입니다.
1. NEBNext UltraExpress™ DNA Library Prep Kit
Single-protocol로 반응 설정 시간이 단축되고, 간소화된 작업 흐름으로 라이브러리 준비 속도가 향상 (manualE3325)
- 10 – 200 ng pre-sheared DNA를 사용하여 2시간 이내에 높은 수율, 고품질 라이브러리를 생성
- Adaptor ligation 후 clean up 과정이 없어 시간이 단축되고, 플라스틱/소모품 사용 감소
- GC-rich, low quality, 또는 low input samples은 기존 제품인 NEBNext Ultra™ II DNA Library Prep Kit (#E7645/E7103) 사용을 권장
Figure 1. The NEBNext UltraExpress™ DNA Library Prep Kit provides robust library yields over a wide input range
Libraries were prepared from 10, 50, 100 or 200 ng of Human NA19240 genomic DNA (Coriell Institute for Medical Research) using the same adaptor amount and 8 PCR cycles. Yields exceeded the minimum requirement (40 ng) for a single Ilumina® NovaSeq® 6000 run to achieve whole genome sequencing with at least 30X coverage.
Q&A
The NEBNext UltraExpressDNA Library Prep Kit is compatible with high-quality human genomic DNA, cell-free DNA (cfDNA) and DNA from other species (e.g., microbial, plant, etc.).
We do not recommend this kit for use with FFPE DNA. For FFPE DNA, consider the NEBNext UltraShear FFPE DNA Library Prep Kit (NEB #E6655).
The NEBNext UltraExpress DNA Library Prep Kit has been optimized for streamlined library prep processing with one adaptor concentration and one PCR cycling condition used for all samples within the 10 – 200 ng input range.
NEBNext Ultra II DNA Library Prep Kit has more customized conditions based on DNA input amount, more steps and cleanups, a longer workflow time, and a broader input range recommendation (500 pg – 1 µg).
Both kits produce high yields of high-quality libraries, but we recommend choosing NEBNext UltraExpress DNA for all of your standard samples within the stated input amount range, where speed is paramount. We recommend choosing NEBNext Ultra II DNA for more complex and precious samples (e.g., GC-rich, low quality, or low input).
No, we recommend using the SPRIselect® Reagent Kit (Beckman Coulter, Inc. #B23317) or AMPure® XP Beads (Beckman Coulter, Inc. #A63881).
The NEBNext UltraExpress Library Prep kits are compatible with both the NEBNext truncated, non-indexed adaptor, supplied with the NEBNext index primer sets and the full-length unique dual index (UDI) unique molecular identifier (UMI) adaptors. For a complete list of all compatible oligos, please see the NEBNext Multiplex Oligos Selection Chart.
2. NEBNext UltraExpress™ RNA Library Prep Kit
Single-protocol로 반응 설정 시간이 단축되고, 간소화된 작업 흐름으로 라이브러리 준비 속도가 향상 (manualE3330)
- Fast workflow (~3 hours): mRNA enrichment 또는 rRNA depletion 과정을 같이 진행해도 하루 만에 고품질 RNA 라이브러리를 생성할 수 있음
- Adaptor ligation 후 clean up 과정이 없어 시간이 단축되고, 플라스틱/소모품 사용 감소
- Automation friendly
- Low quality, 또는 low input samples은 기존 제품인 NEBNext Ultra II Directional RNA Library Prep Kit (#E7760/E7765) 사용을 권장
Figure 2. The NEBNext UltraExpress™ RNA Library Prep Kit produces high library yields across a range of inputs
A. Poly(A)-containing mRNA was isolated from Universal Human Reference RNA (UHRR) (Agilent®), using the NEBNext® Poly(A) mRNA Magnetic Isolation Module (NEB #E7490). Libraries were prepared using the NEBNext UltraExpress™ RNA Library Prep Kit, Kapa mRNA HyperPrep® Kit or Illumina® Stranded mRNA Kit. The NEBNext UltraExpress RNA Library Prep Kit was used with a single adaptor dilution (50X) and 12 PCR cycles for all inputs, or with customized adaptor dilutions (20X for 76–250ng, 100X for 25–75 ng) and PCR cycle numbers (listed in figure).
B. Ribosomal RNA (rRNA) was depleted from UHRR, and libraries were prepared using the UltraExpress RNA Library Prep Kit (preceded by the NEBNext rRNA Depletion Kit (Human/Mouse/Rat – NEB #E7400), Kapa HyperPrep Kit with RiboErase, or Illumina Stranded Total RNA Library Prep Kit with Ribo-Zero® Plus. The NEBNext UltraExpress RNA Library Prep Kit was used with a single adaptor dilution (50X) and 11 PCR cycles for all inputs, or with customized adaptor dilutions (20X for 76–250 ng, 100X for 25–75 ng) and PCR cycle numbers (listed in figure).
The total RNA input amount and number of PCR cycles are indicated. Library yields calculated from an average of three replicates are shown with error bars indicating the standard deviation between replicates.
Q&A
The NEBNext UltraExpress RNA Library Prep Kit has been optimized for streamlined library prep processing with one adaptor concentration and one PCR cycling condition used for all samples within the starting amounts of 25-250 ng of total RNA. The NEBNext Ultra II Directional RNA Library Prep Kit has more customized conditions based on input amount, more steps and cleanups, a longer workflow time, and a broader input range recommendation (10 ng – 1 µg total RNA).
Both kits produce high yields of high-quality libraries, but we recommend choosing NEBNext UltraExpress RNA for all of your standard samples within the stated input amount range, where speed is paramount. We recommend choosing NEBNext Ultra II Directional RNA for more complex and precious samples.
The NEBNext UltraExpress Library Prep kits are compatible with both the NEBNext truncated, non-indexed adaptor, supplied with the NEBNext index primer sets and the full-length unique dual index (UDI) unique molecular identifier (UMI) adaptors. For a complete list of all compatible oligos, please see the NEBNext Multiplex Oligos Selection Chart.
The NEBNext UltraExpress RNA Library Prep Kit has been extensively tested using Universal Human Reference RNA, bacterial RNA mix, and RNA extracted from human whole blood, but it is expected to be compatible with a range of sample types with good quality RNA. While successful libraries have been produced using low-quality samples, optimizing adaptor concentration and PCR cycles is necessary to achieve good quality libraries. We do not recommend using <100 ng of low-quality total RNA (e.g., from FFPE samples) with the NEBNext UltraExpress RNA Library Prep Kit. For highly fragmented RNA samples and low-input, low-quality samples, we recommend using the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (NEB #E7760/E7765).
No. For poly(A) mRNA isolation, we recommend the NEBNext® Poly(A) mRNA Magnetic Isolation Module (NEB #E7490). For rRNA Depletion, we recommend any of our RNA depletion kits (e.g., NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat)) (NEB #E7400/E7405).
NEBNext Selector Tool을 사용하시면 sequencing platform, sample type에 맞춰 권장하는 NEB Next generation sequencing (NGS) 제품을 선택하실 수 있습니다.
