MicroBiology

MiRNA stands for micro RNA and it is abbreviated as miRNA. The miRNA Labeling and detection kit is a unique and a new method developed to label as well as detect the mature miRNA and the small miRNA’s from the entire splinted ligation RNA. This kit uses a proprietary labeling technique that offers exceptional sensitivity to detect the endogenous miRNA. You can now label any sample of RNA such as amplified sense RNA, enriched low molecular weight RNA, degraded RNA and total RNA with the help of this method.

The miRNA Labeling Kit is designed especially in order to enable the labeling of the RNA’s and its molecules for the purpose of microarray analysis with the help of a very simple and a vigorous labeling technique. In this strategy, the RNA molecules are labeled directly, thus offering you a highly selective and a labeling technique for plants, human beings and animals that requires no enrichment at all.

The miRNA Labeling Kit contains the following components:

Clean up columns, positive control, OptiKinase, detection Oligo, clean up mix, 10X OptiKinase Reaction Buffer, Gel Loading Dye, RNase-Free water, a Protocol Card and a Long Protocol Booklet.

Using this technique, one can achieve specificity upto 95%. The labeling method is uniform and extracts reproducible outcome with CV of less than 15% between the demonstrations and less than three logs of active range on most of the scanners like Agilent, Axon, etc.

Advantages of using the miRNA Labeling Kits:

The process is very fast. The direct chemical labeling and capturing of the miRNA is complete in just two hours. Thus, it saves valuable time and the chances of experimental error are at its least. The add-and-incubate reactions take place very simply.The kit can be used flexibly to detect a number of small RNA’s. The linear detection range can be measured quantitatively. It is between the ranges of 0.2 to 20 femtomoles. The miRNA can be detected in as less as 50 ng or even less of the total RNA. Thus, we can say that using the miRNA labeling kit to measure the RNA is a very sensitive specific, fast, flexible and a convenient method.

The miRNA Labeling kit is a four-step process that helps you to label the miRNA directly from the nanogram to microgram quantities. The first step takes about 40 minutes wherein the detection of Oligo preparation takes place. Here the unincorporated nucleotide is taken out with the Clean up column. The second step is capturing the micro RNA that takes about 15 minutes.

MiRNA Ligation is the next step in the process that is completed within 75 minutes. The captured miRNA is ligated and the non-ligated detection Oligo is removed using the Clean up Mix. And the last step is detecting the RNA that required the maximum amount of time for its completion, is time period ranges from three hours to overnight. In this step, the ligated miRNA is separated on a polyacrylamide gel. You can observe this using a phosporimaging system or an X-ray film.