MicroBiology

mRNA refers to messenger RNA. The gene expression studies based on the microarrays enable the simultaneous investigation of the relative amount of messenger RNA for a number of genes with the help of the fluorescently labeled nucleic acid targets. The most of the universal methods makes use of the enzymatic techniques like oligo-dT primed reverse transcription for producing the DNA. These labeling methods have found to have some shortcomings such as laborious protocols, sequence bias, introduced labeling, inability to perceive small alterations in the expression levels and very high experiment-to-experiment variability.

In such cases, a novel and an innovative method of labeling is used. This method makes use of the platinum linked cyanine dyes to chemically and directly label the mRNA from a very little amount of the total RNA. The directly labeled mRNA yields unbiased and highly accurate data of the differential gene expression. The gene expression data that is perceived by using the mRNA labeling method is highly precise without any errors at all. There is no labeling bias and we see a dynamic range over a number of orders of magnitude. This method boasts the highest accuracy levels in identifying the differentially expressed genes. It also cuts down the requirement to run several replicate assays. Thus, minute changes in the expression of genes is now possible to get detected on a large-scale gene expression profile assays with the help of such an easier, quicker as well as simpler procedure.

Nevertheless, the use of the random primers may outwit some of these issues along with the enzymatic labeling techniques, but still we can say that the mRNA Labeling system is quite beneficial. It is an innovative labeling technique that makes use of chemical reagent to label the mRNA directly with the fluorescent dyes. This mRNA is labeled in the entire RNA mixture in a single step non-enzymatic reaction; thereby reducing the errors that have been occurred in the previous methods because of multiple steps. The mRNA thus formed, is free from any labeling bias and it produces reproducible expression profile of the gene and also enables a great amount of accuracy in identifying the differentially expressed genes.

mRNA Labeling System- there are some reagents to be used to prepare the first strand of the DNA for microarray Synthesis. These reagents are highly efficient and full length. Random primers and Oligo dT are used for DNA synthesis. Adequate reagents are provided for ten labeling reactions and they are supplied with the control templates for the Quality Check purpose. This mRNA Labeling system provides the reagents for reverse transcription and for labeling the cDNA from the mRNA templates. The labeled cDNA is, thus used for northern blot applications, FISH or as probes for the microarrays. This mRNA labeling system is stored at a temperature of about 20 degrees C and it remains stable for atleast 6 months.