RNA sequencing (RNA-seq)

The recent development of Next Generation Sequencing enables us to sequence the transcriptome in a certain tissue at a specific developmental time point. RNA-seq reveals a snapshot of the RNAs present and even allows us to quantify the RNAs to determine levels of gene expression.

The instability of RNA molecules makes it necessary to reverse transcribe the transcripts into DNA for sequencing.

To sequence the mRNA we need to perform the following steps:

  1. Isolate the RNA.

  2. Purify mRNA.

  3. Reverse transcribe RNA to cDNA.

  4. DNA modifications for NGS.

  5. Cluster generation for NGS.

  6. NGS.