NEBNext® Small RNA Library Prep Set for Illumina® (Multiplex Compatible)
Note that index primers for multiplexing are not included in this kit. This means that libraries created with this kit cannot be pooled and run together. If index primers are required, kits NEB #E7300, NEB #E7580 or NEB #E7560 should be ordered instead.
The unique workflow of the NEBNext® Small RNA library prep kits addresses the challenge of minimization of adaptor-dimers while achieving production of high-yield, diverse multiplex libraries in a simple protocol.
Please note: index primers for multiplexing are not included in this kit. This version of the kit is intended for use with home brew primers. See FAQ for more information on how to design indexing i7 primer.
Minimized adaptor-dimer contamination
High yields
Input RNA can be Total RNA
Suitable for methylated small RNAs (e.g., piRNAs) as well as unmethylated small RNAs
The NEBNext Multiplex Small RNA Library Prep Kit for Illumina (Index Primers 1-48) contains the adaptors, primers, enzymes and buffers required to convert small RNAs into indexed libraries for next-generation sequencing on the Illumina platform. The novel workflow has been optimized to minimize adaptor-dimers, while producing high-yield, high-diversity libraries.
Please note that this kit, the NEBNextSmall RNA Library Prep Set for Illumina (Multiplex Compatible), enables use with your own barcode system. This kit contains adaptors but does not include multiplex primers. If index primers are required, kits NEB #E7300, NEB #E7580 or NEB #E7560 should be ordered instead.
Each kit component must pass rigorous quality control standards, and each set of reagents is functionally validated together by construction and sequencing of a small RNA library on the Illumina sequencing platform.
The lots provided are managed separately and qualified by additional functional validation. Individual reagents undergo standard enzyme activity and quality control assays, and also meet stringent criteria in the additional quality controls listed on each individual component page
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
E7330S
-20
NEBNext 3´ Ligation Reaction Buffer
E7301AVIAL
-20
1 x 0.24 ml
Not Applicable
Murine RNase Inhibitor
E7308AVIAL
-20
1 x 0.024 ml
Not Applicable
NEBNext 5´ Ligation Reaction Buffer
E7304AVIAL
-20
1 x 0.024 ml
Not Applicable
NEBNext 5´ Ligation Enzyme Mix
E7305AVIAL
-20
1 x 0.06 ml
Not Applicable
LongAmp™ Taq 2X Master Mix
E7309AVIAL
-20
1 x 1.2 ml
Not Applicable
Gel Loading Dye, Blue
E6138AVIAL
-20
1 x 0.2 ml
6 X
Linear Acrylamide
E7325AVIAL
-20
1 x 0.048 ml
Not Applicable
Nuclease-free Water
E7327AVIAL
-20
1 x 5 ml
Not Applicable
Quick-Load pBR322 DNA-MspI Digest
E7323AVIAL
-20
1 x 0.24 ml
Not Applicable
DNA Gel Elution Buffer
E7324AVIAL
-20
1 x 12 ml
Not Applicable
TE Buffer
E7326AVIAL
-20
1 x 0.48 ml
Not Applicable
NEBNext Index 1 Primer for Illumina
E7329AVIAL
-20
1 x 0.12 ml
10 µM
NEBNext 3´ SR Adaptor for Illumina
E7332AVIAL
-20
1 x 0.024 ml
Not Applicable
NEBNext SR RT Primer for Illumina
E7333AVIAL
-20
1 x 0.024 ml
Not Applicable
NEBNext SR Primer for Illumina
E7310AVIAL
-20
1 x 0.06 ml
Not Applicable
NEBNext 5´ SR Adaptor for Illumina
E7328AVIAL
-20
1 x 1,350 pmol
Not Applicable
NEBNext First Strand Synthesis Reaction Buffer
E7334AVIAL
-20
1 x 0.192 ml
Not Applicable
ProtoScript II Reverse Transcriptase
E7355AVIAL
-20
1 x 0.024 ml
Not Applicable
NEBNext 3´ Ligation Enzyme Mix
E7288AVIAL
-20
1 x 0.072 ml
Not Applicable
E7330L
-20
NEBNext 3´ Ligation Reaction Buffer
E7301AAVIAL
-20
1 x 0.96 ml
Not Applicable
Murine RNase Inhibitor
E7308AAVIAL
-20
1 x 0.096 ml
Not Applicable
NEBNext 5´ Ligation Reaction Buffer
E7304AAVIAL
-20
1 x 0.096 ml
Not Applicable
NEBNext 5´ Ligation Enzyme Mix
E7305AAVIAL
-20
1 x 0.24 ml
Not Applicable
LongAmp™ Taq 2X Master Mix
E7309AAVIAL
-20
4 x 1.2 ml
Not Applicable
Gel Loading Dye, Blue
E6138AAVIAL
-20
1 x 1 ml
6 X
Linear Acrylamide
E7325AAVIAL
-20
1 x 0.192 ml
Not Applicable
Nuclease-free Water
E7327AAVIAL
-20
1 x 20 ml
Not Applicable
Quick-Load pBR322 DNA-MspI Digest
E7323AAVIAL
-20
1 x 0.96 ml
Not Applicable
DNA Gel Elution Buffer
E7324AAVIAL
-20
1 x 48 ml
Not Applicable
TE Buffer
E7326AAVIAL
-20
1 x 1.92 ml
Not Applicable
NEBNext Index 1 Primer for Illumina
E7329AAVIAL
-20
1 x 0.48 ml
10 µM
NEBNext 3´ SR Adaptor for Illumina
E7332AAVIAL
-20
1 x 0.096 ml
Not Applicable
NEBNext SR RT Primer for Illumina
E7333AAVIAL
-20
1 x 0.096 ml
Not Applicable
NEBNext SR Primer for Illumina
E7310AAVIAL
-20
1 x 0.24 ml
Not Applicable
NEBNext 5´ SR Adaptor for Illumina
E7328AVIAL
-20
1 x 1,350 pmol
Not Applicable
NEBNext First Strand Synthesis Reaction Buffer
E7334AAVIAL
-20
1 x 0.768 ml
Not Applicable
ProtoScript II Reverse Transcriptase
E7355AAVIAL
-20
1 x 0.096 ml
Not Applicable
NEBNext 3´ Ligation Enzyme Mix
E7288AAVIAL
-20
1 x 0.288 ml
Not Applicable
Properties & Usage
Materials Required but not Supplied
3 M Sodium Acetate, pH 5.5
100% Ethanol
80% Ethanol
Corning®, Costar®, Spin-X® Centrifuge Tube Filters (Cellulose Acetate Filters) (Sigma Aldrich # CLS8162)
Magnetic rack or plate (e.g., NEBNext® Magnetic Separation Rack (NEB #S1515S), Alpaqua ® 96S Super Magnet Plate (#A001322), or equivalent)
Size Selection Materials
for gel size selection:
6% Novex® TBE PAGE gel 1.0 mM
10-well (Life Technologies, Inc. #EC6265BOX)
SYBR® Gold Nucleic Acid Gel Stain (Life Technologies, Inc. #S-11494)
RNase-free Disposable Pellet Pestles® (Kimble Kontes Asset Management, Inc. #749521-1590)
Dry Ice/Methanol Bath or –80°C freezer
for bead selection:
Agencourt® AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
for Pippin Prep™ selection:
3% Agarose Dye Free Gel (Sage Science #CDF 3010)
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Quality, Safety & Legal
Quality Control Assays
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]
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
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