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Universal miRNA Cloning Linker |
 | | | | Universal miRNA Cloning Linker | | | |
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Prices are in US dollars and valid only for US orders.
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Description:
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 10 μl will give a 0.41 μg/μl 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.
Applications:- small RNA cloning (1,2)
- miRNA library construction (1,2)
Storage Conditions
Storage Temperature:
-20°C
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Quality Assurance Statement:
This linker was tested for reactivity with T4 RNA Ligase (1,2). Its identity was confirmed by MALDI-TOF Mass Spectrometry.
References
- Lau et al. (2001) Science, 294, 858-6.
- Pfeffer, S., Lagos-Quintana, M. and Tuschl, T. (2005) Current Protocols in Molecular Biology, (26.4.14) John Wiley & Sons, Inc.
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