In this article, we present an overview of transcription, including requirements of in vitro transcription reactions and a comparison of conventional vs. large scale RNA synthesis.

In vitro transcription (IVT) is a simple procedure that allows for template-directed synthesis of RNA molecules of any sequence from short oligonucleotides to those of several kilobases in μg to mg quantities [1].

In vitro transcription is a simple procedure that allows for template-directed synthesis of RNA molecules of any sequence from short oligonucleotides to those of several kilobases in μg to mg quantities.

In vitro synthesis of single-stranded RNA molecules is a widely used laboratory procedure that is critical to RNA research, as well as to RNA biopharmaceuticals. This technique is versatile in that it allows the researcher to tailor synthesis and introduce modifications to produce a transcript.

In vitro transcription is built upon the central dogma of molecular biology, which states that genetic information flows from DNA to RNA to protein. This process harnesses the natural enzymatic machinery present in cells to produce RNA molecules outside of their native cellular context.

To facilitate experimental analyses, in vitro–synthesized RNAs are often radiolabeled with 32 P (incorporated into specific nucleotides). There are several benefits to using 32 P as an RNA radiolabeling agent. First, compared to other common isotopes, 32 P has the highest emission energy (1.7 MeV), thus increasing the experimental sensitivity.

For many other applications, however, it is crucial that transcription proceeds to the end of the template generating transcripts of all one size (e.g. NPAs, in vitro translation studies, and structural analyses).

We describe a fast, cost-effective and efficient method for in vitro production of tRNA transcripts. T7 RNA polymerase requires a double-stranded DNA promoter in order to initiate transcription; however, elongation does not require a double-stranded DNA template.

Adjust DNA template in DEPC-treated ddH2O to final concentration of 0.5 μg/μl. The DNA clones coming from OpenBiosystems are 0.5 μg/μl, so you do NOT need to adjust. Add in the following order to assemble the transcription reaction.

2010年11月22日 · In vitro transcription is a simple procedure that allows for template-directed synthesis of RNA molecules of any sequence from short oligonucleotides to those of several kilobases in μg to mg quantities.