- Same protocol as the most-commonly used T7/Eberwine in vitro transcription (IVT) kits: No need to change protocol or existing reagents—just add our primers to your reaction, in place of the primers included in your current kit
- Use with commercially-available kits from Ambion, Epicentre, Arcturus, and Amersham
- Has been successfully used for Affymetrix GeneChip® Array analysis
- Study alternative mRNA splicing patterns
- Works with clinical, archived or degraded samples
- Improve yields of amplified RNA with nanogram amounts of starting RNA sample
- Compatible with multiple rounds of T7 IVT amplification
The Full Spectrum™ MultiStart Primers for T7 IVT allow more data to be extracted from your RNA than currently available primers in nearly all commercially-available T7 IVT kits. Just replace the existing T7 primer with the Full Spectrum primers, follow the manufacturer’s protocol, and get more out from your valuable samples.
Simple, Turnkey Solution Eliminates T7 IVT Limitations
Fig 1. Comparison of standard and Full Spectrum T7 IVT priming methods.
Fig 2. Effect of RNA degradation on the standard and Full Spectrum T7 IVT-based RNA amplification methods.
All current RNA amplification methods start by priming first strand cDNA synthesis from the 3’-end poly A tail of mRNA with an oligo-dT primer containing a T7 promoter. This puts a limit on the generation of full length cDNA. Even if the mRNA is highly pure and intact, very often the first strand cDNA is not of full length. This is particularly true with long mRNAs and those with secondary structure, which can impede transcription. This situation becomes worse if the starting mRNA is degraded. As shown in Figures 1 and 2, only the 3’-most portion of the mRNA is present. Unfortunately, RNA obtained from clinical samples is often degraded.
The Full Spectrum™ MultiStart Primer mix initiates first strand cDNA synthesis at multiple points along the mRNA as well as from the poly A tail. This means that the complete mRNA sequence is preserved; even if the mRNA is degraded. Thus, it is now possible to amplify the mRNA while maintaining the complete mRNA sequence.
Kit Information
Full Spectrum™ MultiStart Primers for T7 IVT (provided in a convenient 10X/RT rxn primer mixture)
