Comprehensive Total RNA Seq to Capture Both the mRNA and Small RNA Information in a Single Assay

mRNA sequencing is highly sensitive and an accurate tool for measuring expression across the transcriptome and it provides visibility to undetected changes under different conditions. mRNA-Seq allows researchers to detect both known and novel and it also enables the detection of transcript isoforms, gene fusions, single nucleotide variants, allele-specific gene expression and other features without the limitation of prior knowledge. Small noncoding RNAs also is a part of the whole transcriptome and acts in gene silencing and post-transcriptional regulation of gene expression. Small RNA sequencing can discover novel miRNAs and other small noncoding RNAs without prior sequence or secondary structure information. But as of now an integrated procedure to analyze the mRNA, lncRNA and small RNA's together doesn't exist . Experiments to analyze mRNA and small RNAs separately leads to bias and inconsistency due to sampling , batch and handling variations. We have devised a methodology that can help the researcher to study the entire transcriptome together in a single assay thereby avoiding all the variations and bias. It is more cost effective, more accurate and reliable for concluding results. The target of mRNA degradation or translation repression could be traced from the same sample, which in turn deduces the regulatory role of miRNA.