Materials Required but not Supplied

3 M Sodium Acetate, pH 5.2
100% Ethanol
80% Ethanol
SuperScript® II Reverse Transcriptase (Invitrogen #18064-014)
5X First Strand Buffer (Supplied with SuperScript® II Reverse Transcriptase)
0.1 M DTT (Supplied with SuperScript® II Reverse Transcriptase)
Costar Spin-X centrifuge tube Filters (Cellulose Acetate Filters) (Sigma #CLS8162)
RNase-free Disposable Pellet Pestles (Kontes #74951-1590)
6% Novex TBE PAGE gel, 1.0 mm, 10 well (Invitrogen #EC6265BOX)
SYBR® Gold Nucleic Acid Gel Stain (Invitrogen #S-11494)

Storage Temperature

-20°C

1. Is Index 1 in the NEBNext Small RNA Library Prep Set for SOLiD kit (E6160) compatible with the NEBNext Small RNA Multiplex (1-16) Prep Set for SOLiD kit (E7450)?
2. If my input is <5ug and therefore use 0.5 ul of  3' SR Adapter 3, SR RT Primer 3 and 5' SR Adapter 3, instead of 1.0 ul, my total volumes for each reaction will be short by 0.5 ul. Should I replace that lost volume with water?
3. Why do I add the SR RT Primer 3 before the 5’ ligation instead of right before the reverse transcription step?
4. If I forget to add the 3’ SR RT Primer 3 before the 5’ ligation can I add it before the reverse transcription step and move forward?
5. Can samples be stored longer than 72 hours at -20°C?
6. How many cycles of PCR should I do?
7. If I accidentally move all 30 ul of the 5’ ligation product forward to the reverse transcription mixture will the reaction still work?

1. Library Preparation Using NEBNext® Small RNA Library Prep Set for SOLiD™ (E6160)

• NEBioCalculator

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

• NEBNext® Small RNA Library Prep Set for SOLiD™
• T4 DNA Ligase Reaction Buffer
• Linear Acrylamide
• Murine RNase Inhibitor
• DNA Gel Elution Buffer
• TE Buffer
• Gel Loading Dye, Blue
• NEBNext 3' SR Adaptor 3
• NEBNext SR RT Primer 3
• NEBNext Index 1 Primer for SOLiD
• NEBNext SR Primer R3
• OneTaq Hot Start 2X Master Mix with Standard Buffer
• ProtoScript® II Reverse Transcriptase

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at gbd@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.

Licenses

Notice to Purchaser: Nucleic acid-based aptamers for use with thermophilic DNA polymerases are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. (See Patent Nos. 5,475,096; 5,670,637; 5,696,249; 5,874,557; and 5,693,502). New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use the aptamer-based OneTaq® Hot Start 2X Master Mix with Standard Buffer for RESEARCH PURPOSES ONLY. Commercial use of the aptamer-based OneTaq® Hot Start 2X Master Mix with Standard Buffer requires a license from New England Biolabs, Inc. Please contact busdev@neb.com for more information.