Laboratory Products
New Kit-based Solutions Provide Highly Flexible Microrna Library Creation
Oct 30 2007
Based upon a pre-activated adenylated oligonucleotide linkering method, the miRCat cloning system has been carefully designed to make small RNA library creation easy for all researchers. It consists of three sequential protocols: RNA isolation and enrichment, followed by cloning linker attachment, and ending with an amplification and cloning phase. The system can be used for concatamer cloning or direct ?shotgun' cloning; it also can provide PCR amplicons suitable for use with TOPO TA Cloning® or pGEM® T-Easy cloning vectors.
For cloning small RNAs lacking the 5'-phosphorylated end present in miRNAs, a modified kit - miRCat-33? - has been developed, which uses 5' ligation-independent cloning. This makes it an excellent cloning system for the recently discovered C. elegans 5' tri-phosphorylated microRNAs, which would otherwise be omitted. Furthermore, optional kit components ensure additional versatility with a choice of three different 3' cloning linkers. These provide various restriction site combinations to fit with any preferred cloning processes. Specific primers are also available to convert the resultant small RNA libraries using Roche 454 Life Sciences sequencing systems.
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