Universal miRNA Cloning Linker
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.
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.
Sequence5´ rAppCTGTAGGCACCATCAAT–NH2 3´
- Product Categories:
- miRNA & siRNA Products
- RNA Cloning
- Protocols & Manuals
- Other Tools & Resources
- Quality & Safety
- Legal Information
Ineligible item added to cart
Based on your Freezer Program type, you are trying to add a product to your cart that is either not allowed or not allowed with the existing contents of your cart. Please review and update your order accordingly If you have any questions, please contact Customer Service at email@example.com or 1-800-632-5227 x 8.