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.
Product Information
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
FIGURE 2: The NEBNext RNA Library Prep Kit for MGI 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 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
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
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
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.
This product is related to the following categories:
The following reagents are supplied with this product:
NEB #
Component Name
Component #
Stored at (°C)
Amount
Concentration
E9710S
-20
NEBNext® RNA Fragmentation Buffer
E9618AVIAL
-20
1 x 0.24 ml
Not Applicable
NEBNext First Strand Synthesis Enzyme Mix
E7761AVIAL
-20
1 x 0.048 ml
Not Applicable
NEBNext Strand Specificity Reagent
E7766AVIAL
-20
1 x 0.192 ml
Not Applicable
NEBNext Second Strand Synthesis Enzyme Mix
E7425AVIAL
-20
1 x 0.096 ml
Not Applicable
NEBNext Second Strand Synthesis Reaction Buffer with dUTP Mix
E7426AVIAL
-20
1 x 0.192 ml
Not Applicable
NEBNext® End Prep Reaction Buffer
E9611AVIAL
-20
1 x 0.168 ml
Not Applicable
NEBNext® End Prep Enzyme Mix
E9612AVIAL
-20
1 x 0.072 ml
Not Applicable
NEBNext® Ligation Master Mix
E9613AVIAL
-20
1 x 0.72 ml
Not Applicable
NEBNext® Ligation Enhancer
E7374AVIAL
-20
1 x 0.024 ml
Not Applicable
NEBNext USER® Enzyme
E9619AVIAL
-20
1 x 0.072 ml
Not Applicable
NEBNext® MSTC™ High Yield Master Mix
E9615AVIAL
-20
1 x 0.6 ml
Not Applicable
(0.1X) TE Buffer
E7763AVIAL
-20
1 x 2.78 ml
Not Applicable
Nuclease-free Water
E7764AVIAL
-20
1 x 1.25 ml
Not Applicable
E9710L
-20
NEBNext® RNA Fragmentation Buffer
E9618AAVIAL
-20
1 x 0.96 ml
Not Applicable
NEBNext First Strand Synthesis Enzyme Mix
E7761AAVIAL
-20
1 x 0.192 ml
Not Applicable
NEBNext Strand Specificity Reagent
E7766AAVIAL
-20
1 x 0.768 ml
Not Applicable
NEBNext Second Strand Synthesis Enzyme Mix
E7425AAVIAL
-20
1 x 0.384 ml
Not Applicable
NEBNext Second Strand Synthesis Reaction Buffer with dUTP Mix
E7426AAVIAL
-20
1 x 0.768 ml
Not Applicable
NEBNext® End Prep Reaction Buffer
E9611AAVIAL
-20
1 x 0.672 ml
Not Applicable
NEBNext® End Prep Enzyme Mix
E9612AAVIAL
-20
1 x 0.288 ml
Not Applicable
NEBNext® Ligation Master Mix
E9613AAVIAL
-20
3 x 0.96 ml
Not Applicable
NEBNext® Ligation Enhancer
E7374AAVIAL
-20
1 x 0.096 ml
Not Applicable
NEBNext USER® Enzyme
E9619AAVIAL
-20
1 x 0.288 ml
Not Applicable
NEBNext® MSTC™ High Yield Master Mix
E9615AAVIAL
-20
2 x 1.2 ml
Not Applicable
(0.1X) TE Buffer
E7763AAVIAL
-20
1 x 11.5 ml
Not Applicable
Nuclease-free Water
E7764AAVIAL
-20
1 x 5.7 ml
Not Applicable
Properties & Usage
Materials Required but not Supplied
NEBNext Multiplex Oligos for MGI (Dual Index Primer Pairs Set 1) (#E9725)
NEBNext Circularization Module for MGI (#E9720)
SPRIselect™ Reagent Kit (Beckman Coulter®, Inc. #B23317) or AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
Magnetic Rack (NEB S1515S, Alpaqua® cat. #A001322, or equivalent)
80% Ethanol (freshly prepared)
Thermal cycler
DNase-, RNase-free PCR strip tubes, for example TempAssure® PCR flex-free 8-tube strips (USA Scientific® #1402-4708)
Bioanalyzer® or TapeStation® (Agilent® Technologies, Inc.) and associated reagents and consumables
For use with NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490):
1.5 ml microcentrifuge tube and NEB #S1506 Magnet stand or equivalent (for washing beads only)
For use with NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400) and other NEBNext rRNA depletion kits that do not include beads (NEB #E7750, E7850, E78675):
Agencourt® RNAClean® XP Beads (Beckman Coulter, Inc. #A63987)
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications & Change Notifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
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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