DNA Sequence Library for External RNA Controls

This Standard Reference Material (SRM) is intended for use as a template for ribonucleic acid (RNA) control synthesis using in vitro transcription (IVT).

These RNA controls are designed to be used as external, or “spike-in”, controls to support confidence in gene expression assays by providing quantitative assessment of the technical performance of a gene expression measurement.

A unit of the SRM contains 96 different 0.5 mL polypropylene tubes, with approximately 10 µg, as measured by absorbance (A260), of dehydrated plasmid deoxyribonucleic acid (DNA) in each. 

Each tube contains plasmid DNA with a unique template sequence for a different external RNA control. These controls were developed in cooperation with the External RNA Controls Consortium (ERCC). These templates can be readily used to make RNA controls (see Figure 1). Depending on the strand transcribed, the controls will mimic either “sense” or “anti-sense” eukaryotic messenger RNA (mRNA).

When used to make sense of control RNA, each control will have a nominal 24 nucleotide (nt) polyadenylated (polyA) segment at the 3′ end.

The DNA templates and RNA transcripts produced from them are annotated and described in Figures 2 and 3.

The control sequences range from 273 nt to 2022 nt, with two population distributions of GC fraction(1), one centered at 35 % GC and another at 47 %.

Manufacturer: NIST ( National Institute of Standards and Technology)