Linker-1 is the original modban sequence employed by Lau and Bartel in 2001 and contains a Ban-I restriction site. Linker-2 contains Ava-I and Sty-I restriction sites. Linker-3 contains EcoRI and Msp-I restriction sites and was adapted from Pfeffer and Tuschl. All three linkers are modified with a 3′-terminal dideoxy-C (ddC) base to prevent self ligation.
Experiments have shown miRNA linker performance can vary depending on the RNA source. For this reason, the miRNA Cloning Linker Pack provides small samples (1 nmole) of each of the three linkers and is useful for optimizing methods before using precious RNA samples in large-scale library construction.
Linker oligonucleotides are provided lyophilized and are ready for use in cloning; just resuspend at the desired concentration and add to your ligation mix (using T4 RNA Ligase without ATP). Use of this reagent can improve cloning efficiency of miRNAs, which have a 5′-phosphate and will circularize if attachment of linkers is attempted using RNA Ligase in the presence of ATP.
View microCache 3′ Cloning Linkers on Integrated DNA Technologies’s website