초록 ▼

This invention relates to the use of promoters for ribonucleic acid amplification and other genetic manipulations. Processes are provided wherein complementary deoxyribonucleic acid (cDNA) is synthesized from a ribonucleic acid (RNA) sequence using a complementary primer linked to an RNA polymerase

This invention relates to the use of promoters for ribonucleic acid amplification and other genetic manipulations. Processes are provided wherein complementary deoxyribonucleic acid (cDNA) is synthesized from a ribonucleic acid (RNA) sequence using a complementary primer linked to an RNA polymerase promoter region complement and then anti-sense RNA (aRNA) is transcribed from the cDNA by introducing an RNA polymerase capable of binding to the promoter region. Additional processes using the resulting aRNA are also described.

대표청구항 ▼

[ We claim:] [1.] A gene expression library derived from a cell or tissue sample, comprising two or more specific nucleic acid messages of various abundances, whose levels of representation relative to other messages within a population reflect the physiologic state of the sample, thereby permitting

[ We claim:] [1.] A gene expression library derived from a cell or tissue sample, comprising two or more specific nucleic acid messages of various abundances, whose levels of representation relative to other messages within a population reflect the physiologic state of the sample, thereby permitting diagnosis of a disease or condition, wherein said library is prepared by the following steps:(a) adding a primer complex to a population of messenger RNAs (mRNAs) from said cell or cell population, said primer complex comprising:(i) a primer sequence complementary to a plurality of said population of mRNAs of said cell or cell population, and(ii) a promoter sequence in antisense orientation, wherein said primer complex hybridizes to said plurality population of mRNAs;(b) synthesizing double-stranded complementary deoxyribonucleic acid (cDNA) by(i) extending said primer complex to form a first cDNA strand, and(ii) synthesizing a second cDNA strand complementary to said first cDNA strand without using an exogenous primer, wherein said second cDNA strand comprises said promoter sequence in sense orientation;(c) linearly transcribing multiple copies of double-stranded cDNA into antisense RNAs (aRNAs) initiated from the promoter region of the primer complex; and(d) quantitating specific aRNAs corresponding to specific mRNAs, rein a plurality of said aRNAs represents an expression spectrum in said cell or cell population.