Abstract: Methods for determining the amount of an RNA transcript in a test sample are disclosed, utilizing a reference sample comprising a known amount of a reference nucleic acid, labeled with a first strand 3′ cDNA primer and a first strand 5′ cDNA primer comprising a reference specificity determining box; and a test sample comprising an amount of test RNA containing an RNA transcript of interest, the test RNA being labeled with the first strand 3′ cDNA primer and a first strand 5′ cDNA primer comprising a test specificity determining box. The reference sample and the test sample are mixed and subjected to polymerase chain reaction amplification conditions, followed by division of the amplified, mixed sample and continued amplification of the divided sample to produce nucleic acids containing amplified reference nucleic acid or amplified cDNA of the test RNA, from which cRNA can be generated by in vitro transcription.

Abstract: Methods for determining the amount of an RNA transcript in a test sample are disclosed, utilizing a reference sample comprising a known amount of a reference nucleic acid, labeled with a first strand 3′ cDNA primer and a first strand 5′ cDNA primer comprising a reference specificity determining box; and a test sample comprising an amount of test RNA containing an RNA transcript of interest, the test RNA being labeled with the first strand 3′ cDNA primer and a first strand 5′ cDNA primer comprising a test specificity determining box. The reference sample and the test sample are mixed and subjected to polymerase chain reaction amplification conditions, followed by division of the amplified, mixed sample and continued amplification of the divided sample to produce nucleic acids containing amplified reference nucleic acid or amplified cDNA of the test RNA, from which cRNA can be generated by in vitro transcription.

Abstract: The present invention relates to the discovery of a recombinant vector into which preselected DNA modifications can be readily inserted. Digestion of the vector with a dual cleavage restriction enzyme forms insertion sites which allow the directed placement of an insertion cassette comprised of a double stranded modification fragment containing a preselected sequence modification linked to a pair of single-stranded overhangs with DNA sequences complementary to the DNA sequences of the insertion sites formed in the isolated DNA of the vector. Methods of producing recombinant vectors, and methods of utilizing the vectors to create cell libraries, to identify snRNAs which suppress expression of transcription products, to suppress expression of transcription products and to deliver antisense targeting sequences are also within the scope of the invention.