Patent ID: 8614071
Filing Date: 2013-12-24
Classification: C12Q

Abstract:
1. A method of selective amplification of a messenger RNA (mRNA) target in the presence of corresponding genomic DNA contaminant comprising the steps of: a. providing a sample that comprises said mRNA target and said corresponding genomic DNA contaminant b. hybridizing a first oligonucleotide to said mRNA target and performing RNA-directed DNA synthesis using at least one enzyme capable of RNA-directed synthesis, wherein said first oligonucleotide: i. comprises at least one nucleotide with a base covalently modified at the exocyclic amino group, ii. is at least partially complementary to said mRNA target and to said corresponding genomic DNA contaminant, and, iii. spans an exon-exon junction in the target; c. amplifying the product of step b) using said first oligonucleotide and a second oligonucleotide with at least one enzyme capable of DNA-directed DNA synthesis; wherein said second oligonucleotide is at least partially complementary to said mRNA target and to said corresponding genomic DNA contaminant; wherein said mRNA target is selectively amplified over said corresponding genomic DNA contaminant when compared to amplifying with an unmodified first oligonucleotide.