NEBNext^® Multiplex Small RNA Library Prep Kit for Illumina^® (Index Primers 1-48)

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
E7560S     -20       Gel Loading Dye, Blue E6138AAVIAL -20 1 x 1 ml 6 X   NEBNext 3´ Ligation Enzyme Mix E7288AAVIAL -20 1 x 0.288 ml Not Applicable   NEBNext Index 1 Primer for Illumina E7311AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 2 Primer for Illumina E7312AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 3 Primer for Illumina E7313AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 4 Primer for Illumina E7314AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 5 Primer for Illumina E7315AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 7 Primer for Illumina E7317AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 8 Primer for Illumina E7318AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 9 Primer for Illumina E7319AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 10 Primer for Illumina E7320AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 11 Primer for Illumina E7321AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 12 Primer for Illumina E7322AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 6 Primer for Illumina E7316AVIAL -20 1 x 0.01 ml 10 µM   NEBNext® Index 25 Primer for Illumina® E7561AVIAL -20 1 x 0.01 ml 10 µM   NEBNext® Index 27 Primer for Illumina® E7562AVIAL -20 1 x 0.01 ml 10 µM   NEBNext 3´ Ligation Reaction Buffer E7301AAVIAL -20 1 x 0.96 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   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   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 5´ SR Adaptor for Illumina E7328AVIAL -20 1 x 1,350 pmol Not Applicable   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 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 Index 26 Primer for Illumina E7713AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 28 Primer for Illumina E7714AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 29 Primer for Illumina E7715AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 30 Primer for Illumina E7716AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 31 Primer for Illumina E7717AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 32 Primer for Illumina E7718AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 33 Primer for Illumina E7719AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 34 Primer for Illumina E7720AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 35 Primer for Illumina E7721AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 36 Primer for Illumina E7722AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 37 Primer for Illumina E7731AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 38 Primer for Illumina E7732AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 39 Primer for Illumina E7733AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 40 Primer for Illumina E7734AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 41 Primer for Illumina E7735AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 42 Primer for Illumina E7736AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 43 Primer for Illumina E7737AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 44 Primer for Illumina E7738AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 45 Primer for Illumina E7739AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 46 Primer for Illumina E7740AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 47 Primer for Illumina E7741AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 48 Primer for Illumina E7742AVIAL -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 24 Primer for Illumina E7592AVIAL -20 1 x 0.01 ml 10 µM   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 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 18 Primer for Illumina E7586AVIAL -20 1 x 0.01 ml 10 µM   Quick-Load pBR322 DNA-MspI Digest E7323AAVIAL -20 1 x 0.96 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)

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 with the NEBNext^® Small RNA Library prep kit for Illumina compatible with paired-end flow cells 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?
16. Do I have to use custom sequencing primers for my libraries generated with NEBNext^® Small RNA Library prep kit for Illumina?

NEBNext Product Citations Search Tool

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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.

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]

• E7560S_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]

• E7560S_v0_10009711
• E7560S_v0_10014422
• E7560S_v0_10019288
• E7560S_v0_10022570
• E7560S_v0_10033517
• E7560S_v0_10034898
• E7560S_v0_10022124
• E7560S_v1_10042541
• E7560S_v1_10042841
• E7560S_v1_10047658
• E7560S_v1_10049679
• E7560S_v1_10037545
• E7560S_v1_10061906
• E7560S_v1_10062733
• E7560S_v1_10065077
• E7560S_v1_10077492
• E7560S_v1_10086114
• E7560S_v1_10090554
• E7560S_v1_10095643
• E7560S_v1_10101556
• E7560S_v1_10106473
• E7560S_v1_10111113
• E7560S_v1_10093789
• E7560S_v1_10122801
• E7560S_v1_10113371
• E7560S_v1_10150201
• E7560S_v1_10161950
• E7560S_v1_10166614
• E7560S_v1_10168340
• E7560S_v1_10171978
• E7560S_v1_10136724
• E7560S_v1_10182215
• E7560S_v1_10186418
• E7560S_v1_10196487
• E7560S_v1_10172748
• E7560S_v1_10193959
• E7560S_v1_10224288
• E7560S_v1_10246351

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

• NEBNext^® Multiplex Small RNA Library Prep Kit for Illumina^® (Index Primers 1-48)
• Gel Loading Dye, Blue
• NEBNext 3´ Ligation Enzyme Mix
• NEBNext Index 1 Primer for Illumina
• NEBNext Index 2 Primer for Illumina
• NEBNext Index 3 Primer for Illumina
• NEBNext Index 4 Primer for Illumina
• NEBNext Index 5 Primer for Illumina
• NEBNext Index 7 Primer for Illumina
• NEBNext Index 8 Primer for Illumina
• NEBNext Index 9 Primer for Illumina
• NEBNext Index 10 Primer for Illumina
• NEBNext Index 11 Primer for Illumina
• NEBNext Index 12 Primer for Illumina
• NEBNext Index 6 Primer for Illumina
• NEBNext 3´ Ligation Reaction Buffer
• NEBNext 5´ Ligation Reaction Buffer
• NEBNext 5´ Ligation Enzyme Mix
• Murine RNase Inhibitor
• LongAmp™ Taq 2X Master Mix
• NEBNext SR Primer for Illumina
• DNA Gel Elution Buffer
• Linear Acrylamide
• TE Buffer
• Nuclease-free Water
• NEBNext 5´ SR Adaptor for Illumina
• NEBNext 3´ SR Adaptor for Illumina
• NEBNext SR RT Primer for Illumina
• NEBNext First Strand Synthesis Reaction Buffer
• ProtoScript II Reverse Transcriptase
• NEBNext Index 26 Primer for Illumina
• NEBNext Index 28 Primer for Illumina
• NEBNext Index 29 Primer for Illumina
• NEBNext Index 30 Primer for Illumina
• NEBNext Index 31 Primer for Illumina
• NEBNext Index 32 Primer for Illumina
• NEBNext Index 33 Primer for Illumina
• NEBNext Index 34 Primer for Illumina
• NEBNext Index 35 Primer for Illumina
• NEBNext Index 36 Primer for Illumina
• NEBNext Index 37 Primer for Illumina
• NEBNext Index 38 Primer for Illumina
• NEBNext Index 39 Primer for Illumina
• NEBNext Index 40 Primer for Illumina
• NEBNext Index 41 Primer for Illumina
• NEBNext Index 42 Primer for Illumina
• NEBNext Index 43 Primer for Illumina
• NEBNext Index 44 Primer for Illumina
• NEBNext Index 45 Primer for Illumina
• NEBNext Index 46 Primer for Illumina
• NEBNext Index 47 Primer for Illumina
• NEBNext Index 48 Primer for Illumina
• NEBNext Index 16 Primer for Illumina
• NEBNext Index 17 Primer for Illumina
• NEBNext Index 24 Primer for Illumina
• NEBNext Index 13 Primer for Illumina
• NEBNext Index 14 Primer for Illumina
• NEBNext Index 15 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 18 Primer for Illumina
• Quick-Load pBR322 DNA-MspI Digest

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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.