Introduction
T7 RNA Synthesis Kit is designed for
in vitro transcription of large amounts of RNA, provides with T7 RNA polymerase mix and four individually available nucleotides. The kit can also be used to synthesise capped RNA by adding cap analogues; or to synthesise modified RNA by replacing with modified UTP according to experimental requirements. The kit contains LiCl Solution for removing free nucleotides, enzymes and most of the template from the transcription product to obtain coarsely purified RNA.
Advantage
- Yields 150~200 μg of RNA from 1 μg of DNA template.
- Optimized transcription buffer: Requires less T7 RNA polymerase for maximum RNA yield compared to other competitors.
Application
Synthesis of large amounts of various RNAs, including but not limited to:
- mRNA for vaccine or gene expression
- precursors of circular RNA
- labeled RNA probes
- Modified RNA: prepared by admixing modified NTP (e.g., aminoallyl-, biotin-, fluorescein-, digoxin-NTP)
Performance
RNA yield
Fig. 1 Yield for different reaction durations
1 μg of 2600 bp DNA template was added to 20 μL of the reaction system and incubated at 37 °C for 30~120 min. The IVT products were purified with LiCl and analysed by NanoDrop™ to determine the amount of product for different reaction durations (Figure 1.A); and an aliquot of the product was electrophoresed on a 1 % agarose gel (Figure 1.B).
Competitor Comparison
Fig. 2 Comparison of in vitro transcribed RNA yields
Compare the yield of RNA for in vitro transcription with PC028+, PC035 kit, and a competitor IVT kit (NEB, #E2040). PC028+ represents the substitution of GeneCopoeia’s T7 RNA polymerase for the T7 RNA polymerase in the competitor’s IVT kit. The reaction system was configured using 1 μg of Control Template DNA from the PC035 kit as a template and incubated at 37 degrees for 2 hours. The final products were purified by LiCl and 1 μL of each was taken for electrophoretic analysis (Figure A). And Compared the yields calculated after measuring the concentration using Nanodrop (Figure B). The ssRNA ladder was loaded with 2 μL.
