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Universal miRNA Cloning Linker

Catalog # Concentration Size List Price Quantity Your Price
S1315S 5 µg $255.00
$229.50
Catalog # Size List Price Your Price
S1315S 5 µg $255.00
$229.50
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • This 5´ adenylated, 3´ blocked oligodeoxynucleotide can be used for cloning short RNAs according to the procedure of Bartel (1).
  • RNA ligase recognizes the "activated" adenylated oligo and covalently ligates its 5´ end to the 3´ OH of a second single stranded sequence in the absence of ATP.
  • In a mixture of nucleic acids use of the 5´ adenylated, 3´ blocked oligo with RNA ligase (w/o ATP) results in ligation of the target oligo only.
  • Sequence: 5′ rAppCTGTAGGCACCATCAAT–NH2 3′
Linker/RNA Ligation Protocol: The Universal miRNA cloning linker has a 3´ blocking group (amine) to prevent self ligation, circularization and ligation to RNA at the 5´ end. It also contains the BanI restriction site compatible with the miRNA cloning protocol used by Bartel (1).

For ligation, resuspend the lyophilized oligonucleotide in RNAse-free water to 100 μM stock concentration. (A resuspension volume of 8.6 μl will give a 0.58 μg/μl or 100 μM stock solution). Ligations are carried out with T4 RNA Ligase I in the absence of ATP. Approximately 1 μg is typically used per reaction. See reference 1 for a detailed protocol.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Universal miRNA Cloning Linker S1315SVIAL -20 1 x 5 µg
Application Features
  • small RNA cloning (1,2)
  • miRNA library construction (1,2)

Properties & Usage

Suggested Working Concentration

100μM

Recommended Resuspension Volume

8.6 μl

References
  • Lau et al. (2001). Science. 294, 858-6.
  • Pfeffer, S., Lagos-Quintana, M. and Tuschl, T. (2005). Current Protocols in Molecular Biology, John Wiley & Sons, Inc. (26.4.14).
Quality Control Assay
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
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]
Certificate of Analysis
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]
Safety Data Sheets
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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.

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