Full
Spectrum™ MultiStart Expands T7 IVT RNA Amplification to the Entire
Transcriptome
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 MORE >>
Use with
commercially-available kits from Ambion, Epicentre, Arcturus, and Amersham
Has been successfully
used for Affymetrix GeneChip® Array analysis
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.
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)
Full Spectrum Complete Transcriptome Amplification Kit.
For quantitative PCR (qPCR) applications, we recommend the PCR-based Full
Spectrum Complete Transcriptome Amplification Kit. With all of the same advantages as T7 IVT,
you can generate enough amplified cDNA for qPCR expression analysis of
hundreds of transcripts in fewer steps, with less cost, and in under 3
hours.
