Nuclear Extract in vitro Transcription System, HeLaScribe®

HeLaScribe® Nuclear Extract in vitro Transcription System provides a cell-free system for in vitro transcription of eukaryotic genes.

Performance-tested with cytomegalovirus immediate early gene (CMV) promoter
Source of transcription factors, DNA-binding proteins and RNA processing components

The most well characterised cell-free system for in vitro transcription of eukaryotic genes is derived from HeLa cell nuclei. HeLa nuclear extracts can support accurate transcription initiation by RNA polymerase II and exhibit both basal and regulated patterns of RNA polymerase transcription. These nuclear extracts also are a source for a variety of transcription factors, DNA binding proteins and the enzymatic machinery involved in RNA processing.

The HeLaScribe® Nuclear Extract included in this system is prepared by a modification of the method of Dignam, et al. The system includes all of the necessary components for in vitro transcription as well as a positive control template that provides run-off transcripts from a CMV immediate early promoter.

This system is designed for run-off transcription. Alternatively, transcription products can be analysed by primer extension.

: Complete system (sufficient for 40 reactions) includes 320 U HeLaScribe® Nuclear Extract, in vitro transcription grade; 400 µl HeLa Nuclear Extract 1X Transcription Buffer; 1×15 ml HeLa Extract Stop Solution; 500 µl MgCl₂, 50 mM; 15 µl each of rATP, rCTP, rGTP, rUTP (100 mM); 300 ng HeLa Nuclear Extract Positive Control DNA (CMV); 1 ml Loading Dye; and 1,25 ml nuclease-free water. HeLaScribe® Nuclear Extract, in vitro transcription grade also available separately.