RNA sequencing is one of the most powerful and novel techniques for transcriptomic studies. However, one of the biggest drawbacks is the high cost associated with such type of high-throughput analysis making this technology unavailable to several laboratories worldwide.
Besides the price, since RNAseq is a relatively new technique, some technical issues have been found and improvements should be done in order to obtain more informative data (such as strand specificity).
Hence, this group of investigators have decided to develop a low-cost library construction protocol that is compatible with the current dominate RNA-seq platform provider, illumina Inc, reducing the expenses in 90% as also the bench time required.
The molecular biology field has a wide range of tools available, and now, in the perspective of transcriptomics analysis, there’s a new exciting method called RNA sequencing (RNA-seq).
In this paper the authors review, as the title indicates, the advances, challenges and opportunities of this new technique. Also, they explore the different RNA-seq approaches such as next generation sequencing methods that can be applied, the advantages or limitations of direct or indirect RNA sequencing and finally, important concepts and diagrams that help us to understand the principle behind the method.
Since I am planning to do B-cell ELIspot I decided to write and this way learn about this technique.
B-cell ELIspot was fist described in 1983 by Czerkinsky and collaborators in order enumerate the B-cells secreting antigen-specific antibodies. The principle is very similar to the sandwich ELISA but ELIspot works at a single cell level giving us both quantitative and qualitative information with high sensitivity.
However, the method had suffered some alterations and started to be used for the detection of total antibodies produced by B-cells or to analyse antigen-specific T-cell response. The application of this technique is vast, being used more frequently in the characterization of B-cell response eliceted by infections and vaccination.