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NEBNext® RNA Library Prep Kit for MGI®

Catalog # Concentration Size List Price Quantity Your Price
E9710L 96 reactions $3,856.00
$3,470.40
E9710S 24 reactions $1,070.00
$963.00
Catalog # Size List Price Your Price
E9710L 96 reactions $3,856.00
$3,470.40
E9710S 24 reactions $1,070.00
$963.00
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

The NEBNext RNA 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 directional RNA libraries from a wide range of inputs (10 ng – 1 µg total RNA). The kit can be used with rRNA depletion or poly(A) enrichment protocols, including the Express protocol for the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490), as detailed in the product manual.

The NEBNext RNA Library Prep Kit for MGI 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: 10 ng – 1 µg total RNA
  • Uniform transcript and GC coverage
  • 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.

 

The NEBNext RNA Library Prep Kit for MGI meets the need for reliable high-quality directional RNA library prep for MGI sequencing applications.

The kit incorporates trusted NEBNext master mixed reagents and protocols and allows the use of a wide range of inputs (10 ng – 1 µg total RNA). rRNA depletion or poly(A) enrichment protocols can be used upstream of the kit, and the Express protocol for the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) (see product manual) provides additional time savings.

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 10 ng to 1 µg total RNA
  • Increase the complexity and transcript coverage of your libraries
  • Optimize your time with streamlined workflows, minimized hands-on time, and automation compatibility
  • Rely on robust performance

 

FIGURE 1: NEBNext RNA Library Prep Kit for MGI workflow

E9710 Workflow

FIGURE 2: The NEBNext RNA Library Prep Kit for MGI produces high library yields across a range of inputs

E9710 MGI Yield


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 RNA Library Prep Kit for MGI using the recommendations for adaptor dilution and PCR cycles included in the kit protocol.

B. Ribosomal RNA (rRNA) was depleted from UHRR (NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400), and libraries were prepared using the NEBNext RNA Library Prep Kit for MGI, following the recommendations for adaptor dilution and PCR cycles included in the kit protocol. Library yields calculated from an average of three replicates are shown with error bars indicating the standard deviation between replicates.

FIGURE 3: The NEBNext RNA Library Prep Kit for MGI provides consistent transcript coverage

E7910 Transcript Coverage

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 RNA Library Prep Kit for MGI using the recommendations for adaptor dilution and PCR cycles included in the kit protocol.

B. Ribosomal RNA (rRNA) was depleted from UHRR (NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat), NEB #E7400), and libraries were prepared using the NEBNext RNA Library Prep Kit for MGI, following the recommendations for adaptor dilution and PCR cycles included in the kit protocol.  Libraries were sequenced on an MGI DNBSEQ-G400 (2 x 100 bases). 10 million reads were sampled and mapped to the hg38 reference genome using RNA STAR v2.7.8a and 5´ to 3´ transcript coverage was calculated from the top 1,000 transcripts using the CollectRnaSeqMetrics (Picard) tool v2.18.2.2. Even and consistent coverage was observed across all input amounts.


FIGURE 4: The NEBNext RNA Library Prep Kit for MGI provides high levels of detection for high- and low-abundance transcripts

E7910 High Detection

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 RNA Library Prep Kit for MGI using the recommendations for adaptor dilution and PCR cycles included in the kit protocol.

B. Ribosomal RNA (rRNA) was depleted from UHRR (NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat), NEB #E7400), and libraries were prepared using the NEBNext RNA Library Prep Kit for MGI, following the recommendations for adaptor dilution and PCR cycles included in the kit protocol.  Libraries were sequenced on an MGI DNBSEQ-G400 (2 x 100 bases). 10 million reads were sampled, and average transcript abundance was assessed for transcripts detected with ≤ 10 reads and ≤ 500 reads, respectively. Error bars indicate standard deviation for 3 replicates. Salmon v1.5.1 was used for mapping and quantification of all gencode v38 transcripts and ERCCs. A high level of transcript detection was observed across all input amounts and the two workflows.


FIGURE 5: The NEBNext RNA library prep kit for MGI provides excellent transcript correlation between inputs and replicates

E7910 Transcript Correlation

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 RNA Library Prep Kit for MGI. The NEBNext RNA Library Prep Kit for MGI was used using the recommendations for adaptor dilution and PCR cycles included in the kit protocol.

B. Ribosomal RNA (rRNA) was depleted from UHRR (NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat), NEB #E7400) and libraries were prepared using the NEBNext RNA Library Prep Kit for MGI, following the recommendations for adaptor dilution and PCR cycles included in the kit protocol. Libraries were sequenced on an MGI DNBSEQ-G400 (2 x 100 bases). 10 million reads were sampled and libraries were correlated for transcript expression levels across inputs using Salmon v1.5.1 quantification of all gencode v38 transcripts and ERCCs. Each data point represents a transcript, with the log10 abundance in Read Counts with 1 μg total RNA input on the y-axis compared to a replicate at 1 μg followed by 200 ng, 100 ng and 10 ng on the x-axis. The number of transcripts detected have very high correlation across the different inputs and replicates.

 
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  NEBNext® RNA Fragmentation Buffer E9618AVIAL -20 1 x 0.24 ml
  NEBNext First Strand Synthesis Enzyme Mix E7761AVIAL -20 1 x 0.048 ml
  NEBNext Strand Specificity Reagent E7766AVIAL -20 1 x 0.192 ml
  NEBNext Second Strand Synthesis Enzyme Mix E7425AVIAL -20 1 x 0.096 ml
  NEBNext Second Strand Synthesis Reaction Buffer with dUTP Mix E7426AVIAL -20 1 x 0.192 ml
  NEBNext® End Prep Reaction Buffer E9611AVIAL -20 1 x 0.168 ml
  NEBNext® End Prep Enzyme Mix E9612AVIAL -20 1 x 0.072 ml
  NEBNext® Ligation Master Mix E9613AVIAL -20 1 x 0.72 ml
  NEBNext® Ligation Enhancer E7374AVIAL -20 1 x 0.024 ml
  NEBNext USER® Enzyme E9619AVIAL -20 1 x 0.072 ml
  NEBNext® MSTC™ High Yield Master Mix E9615AVIAL -20 1 x 0.6 ml
  (0.1X) TE Buffer E7763AVIAL -20 1 x 2.78 ml
  Nuclease-free Water E7764AVIAL -20 1 x 1.25 ml
  NEBNext® RNA Fragmentation Buffer E9618AAVIAL -20 1 x 0.96 ml
  NEBNext First Strand Synthesis Enzyme Mix E7761AAVIAL -20 1 x 0.192 ml
  NEBNext Strand Specificity Reagent E7766AAVIAL -20 1 x 0.768 ml
  NEBNext Second Strand Synthesis Enzyme Mix E7425AAVIAL -20 1 x 0.384 ml
  NEBNext Second Strand Synthesis Reaction Buffer with dUTP Mix E7426AAVIAL -20 1 x 0.768 ml
  NEBNext® End Prep Reaction Buffer E9611AAVIAL -20 1 x 0.672 ml
  NEBNext® End Prep Enzyme Mix E9612AAVIAL -20 1 x 0.288 ml
  NEBNext® Ligation Master Mix E9613AAVIAL -20 3 x 0.96 ml
  NEBNext® Ligation Enhancer E7374AAVIAL -20 1 x 0.096 ml
  NEBNext USER® Enzyme E9619AAVIAL -20 1 x 0.288 ml
  NEBNext® MSTC™ High Yield Master Mix E9615AAVIAL -20 2 x 1.2 ml
  (0.1X) TE Buffer E7763AAVIAL -20 1 x 11.5 ml
  Nuclease-free Water E7764AAVIAL -20 1 x 5.7 ml
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