NEBNext^® Multiplex Small RNA Library Prep Set for Illumina^® (Set 2)

Kit Components

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
E7580S     -20       NEBNext 3´ Ligation Reaction Buffer E7301AVIAL -20 1 x 0.24 ml Not Applicable   NEBNext 3´ SR Adaptor for Illumina E7332AVIAL -20 1 x 0.024 ml Not Applicable   NEBNext 5´ SR Adaptor for Illumina E7328AVIAL -20 1 x 1,350 pmol 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   NEBNext SR RT Primer for Illumina E7333AVIAL -20 1 x 0.024 ml 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   Murine RNase Inhibitor E7308AVIAL -20 1 x 0.024 ml Not Applicable   LongAmp™ Taq 2X Master Mix E7309AVIAL -20 1 x 1.2 ml Not Applicable   NEBNext SR Primer for Illumina E7310AVIAL -20 1 x 0.06 ml Not Applicable   Gel Loading Dye, Blue E6138AVIAL -20 1 x 0.2 ml 6 X   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   Linear Acrylamide E7325AVIAL -20 1 x 0.048 ml Not Applicable   TE Buffer E7326AVIAL -20 1 x 0.48 ml Not Applicable   Nuclease-free Water E7327AVIAL -20 1 x 5 ml Not Applicable   NEBNext Index 13 Primer for Illumina E7581AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 14 Primer for Illumina E7582AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 15 Primer for Illumina E7583AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 16 Primer for Illumina E7584AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 17 Primer for Illumina E7585AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 18 Primer for Illumina E7586AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 19 Primer for Illumina E7587AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 20 Primer for Illumina E7588AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 21 Primer for Illumina E7589AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 22 Primer for Illumina E7590AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 23 Primer for Illumina E7591AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 24 Primer for Illumina E7592AVIAL -20 1 x 0.01 ml 10 µM   NEBNext 3´ Ligation Enzyme Mix E7288AVIAL -20 1 x 0.072 ml Not Applicable
E7580L     -20       NEBNext 3´ Ligation Reaction Buffer E7301AAVIAL -20 1 x 0.96 ml Not Applicable   NEBNext 3´ SR Adaptor for Illumina E7332AAVIAL -20 1 x 0.096 ml Not Applicable   NEBNext 5´ SR Adaptor for Illumina E7328AVIAL -20 1 x 1,350 pmol 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   NEBNext SR RT Primer for Illumina E7333AAVIAL -20 1 x 0.096 ml 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   Murine RNase Inhibitor E7308AAVIAL -20 1 x 0.096 ml Not Applicable   LongAmp™ Taq 2X Master Mix E7309AAVIAL -20 4 x 1.2 ml Not Applicable   NEBNext SR Primer for Illumina E7310AAVIAL -20 1 x 0.24 ml Not Applicable   Gel Loading Dye, Blue E6138AAVIAL -20 1 x 1 ml 6 X   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   Linear Acrylamide E7325AAVIAL -20 1 x 0.192 ml Not Applicable   TE Buffer E7326AAVIAL -20 1 x 1.92 ml Not Applicable   Nuclease-free Water E7327AAVIAL -20 1 x 20 ml Not Applicable   NEBNext Index 13 Primer for Illumina E7581AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 14 Primer for Illumina E7582AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 15 Primer for Illumina E7583AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 16 Primer for Illumina E7584AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 17 Primer for Illumina E7585AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 18 Primer for Illumina E7586AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 19 Primer for Illumina E7587AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 20 Primer for Illumina E7588AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 21 Primer for Illumina E7589AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 22 Primer for Illumina E7590AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 23 Primer for Illumina E7591AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 24 Primer for Illumina E7592AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext 3´ Ligation Enzyme Mix E7288AAVIAL -20 1 x 0.288 ml Not Applicable

1. Protocol for use with NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
2. 6A. Quality Control Check and Size Selection using 6% Polyacrylamide Gel  NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
3. 6B. Quality Control Check and Size Selection using Pippin Prep NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
4. 6C. Quality Control Check and Size Selection using AMPure XP Beads NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)

• Automated size selection of NEBNext^® Small RNA libraries with the Sage Pippin Prep™

1.  Is it possible to prevent abundant, unwanted RNAs to be included in the Small RNA library?
2. Can I use Total RNA to make small RNA libraries or do I have to isolate or enrich the sample for small RNA?
3. Why does the RT primer hybridization occur before 5’adaptor ligation?
4. Do I have to hybridize the RT primer again after 5’ ligation?
5. How are the barcodes introduced in the Multiplex libraries?
6. During size selection on 6% PAGE gel, which bands should I cut out of the gel?
7. Are libraries prepared by this method compatible with paired-end flowcells for cluster generation?
8. Can I use the small RNA sample preparation kit for Directional-RNA sequencing?
9. What is the sequence of the final PCR product?
10. Can I use enriched small RNA instead of total RNA for the NEBNext Small RNA Library Prep for Illumina?
11. What should I do if my Small RNA may not have 5´ Phosphate and 3´ OH groups?
12. How should my NEBNext Small RNA library be trimmed?
13. What should I do if the sample sheet from the NEB.com website is not compatible with the software version I am using?
14. Do you have Sample Sheets for use with the NEBNext Multiplex Small RNA Library Prep for Illumina?
15. Are NEBNext adaptor and primers for Illumina compatible with NEBNext reagents for Illumina library preparation?

NEBNext Product Citations Search Tool

Use this tool to find citations related to NEBNext products. Search by product, keyword or instrument name – use the filters to select specific product areas.

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]

• E7580S_L_v1

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]

• E7580L_v0_10012749
• E7580L_v0_10012794
• E7580S_v0_10013886
• E7580L_v0_10015910
• E7580S_v0_10019812
• E7580L_v0_10027819
• E7580S_v0_10026805
• E7580S_v0_10032731
• E7580L_v0_10029939
• E7580S_v0_10033910
• E7580S_v0_10037241
• E7580S_v1_10043928
• E7580S_v1_10037491
• E7580S_v1_10053194
• E7580L_v1_10056242
• E7580S_v1_10058115
• E7580L_v1_10060362
• E7580L_v1_10060691
• E7580S_v1_10060469
• E7580L_v1_10062765
• E7580S_v1_10063253
• E7580L_v1_10053082
• E7580S_v1_10065073
• E7580L_v1_10073775
• E7580S_v1_10083156
• E7580S_v1_10084686
• E7580L_v1_10089508
• E7580S_v1_10090208
• E7580S_v1_10092532
• E7580S_v1_10095626
• E7580L_v1_10095639
• E7580S_v1_10099994
• E7580S_v1_10066132
• E7580S_v1_10105681
• E7580L_v1_10093736
• E7580S_v1_10111739
• E7580S_v1_10119179
• E7580S_v1_10128025
• E7580S_v1_10135904
• E7580L_v1_10142386
• E7580S_v1_10154243
• E7580S_v1_10157244
• E7580L_v1_10159867
• E7580S_v1_10163566
• E7580L_v1_10136737
• E7580S_v1_10165331
• E7580S_v1_10169990
• E7580S_v1_10174419
• E7580S_v1_10136765
• E7580L_v1_10172769
• E7580S_v1_10182117

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• NEBNext 3´ Ligation Reaction Buffer
• NEBNext 3´ SR Adaptor for Illumina
• NEBNext 5´ SR Adaptor for Illumina
• NEBNext 5´ Ligation Reaction Buffer
• NEBNext 5´ Ligation Enzyme Mix
• NEBNext SR RT Primer for Illumina
• NEBNext First Strand Synthesis Reaction Buffer
• ProtoScript II Reverse Transcriptase
• Murine RNase Inhibitor
• LongAmp™ Taq 2X Master Mix
• NEBNext SR Primer for Illumina
• Gel Loading Dye, Blue
• Quick-Load pBR322 DNA-MspI Digest
• DNA Gel Elution Buffer
• Linear Acrylamide
• TE Buffer
• Nuclease-free Water
• NEBNext Index 13 Primer for Illumina
• NEBNext Index 14 Primer for Illumina
• NEBNext Index 15 Primer for Illumina
• NEBNext Index 16 Primer for Illumina
• NEBNext Index 17 Primer for Illumina
• NEBNext Index 18 Primer for Illumina
• NEBNext Index 19 Primer for Illumina
• NEBNext Index 20 Primer for Illumina
• NEBNext Index 21 Primer for Illumina
• NEBNext Index 22 Primer for Illumina
• NEBNext Index 23 Primer for Illumina
• NEBNext Index 24 Primer for Illumina
• NEBNext 3´ Ligation Enzyme Mix

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 busdev@neb.com.

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.