Absolute quantitation of nucleic acids by rt-pcr
OwnerNormand Allaireinventor
Life of this patent
Estimating the term…
Abstract
A method for obtaining a cRNA for use in generating calibration data, e.g., a standard curve, for absolute quantitation of RNA by RT-PCR is disclosed. The method includes the steps of: providing a synthetic oligonucleotide comprising an amplicon, a promoter sequence located 3′ relative to the amplicon; synthesizing complementary RNA (cRNA) by in vitro transcription of the synthetic oligonucleotide; quantitatively assaying the cRNA by an independent method; and generating calibration data using a known quantity of the cRNA.
Claims24
1-18. (canceled)
A calibrator for absolute quantitation of target RNA by RT-PCR comprising a single cRNA species synthesized by reverse transcription from a synthetic oligonucleotide and quantitatively assayed by an independent method, wherein said synthetic oligonucleotide comprises an amplicon and a promoter sequence located 3′ relative to the amplicon.
Dependencies are inferred from the claim text.