Abstract
Abstract In the presence of messenger or viral RNA containing poly(A) stretches as a template and (dT)10 as a primer, DNA polymerase I from Escherichia coli synthesizes poly(dT) using dTTP as the sole precursor. No synthesis occurs when dATP is present as the sole precursor, although dATP is incorporated into acid-insoluble product in the presence of dTTP. Identical results were observed using the synthetic homopolymeric template poly(rA) with (dT)10 as added primer. Thus, poly(dT) synthesized on poly(A) stretches appears to serve as a template for the synthesis of poly(dA). Poly(dT) which has been synthesized using (dA)n·(dT)10 as template-primer does not allow the incorporation of dATP in the presence of dTTP. We have utilized these observations as a basis for the development of an extremely sensitive and precise technique for the detection of poly(A) stretches in any RNA sample. This technique can also be used in the presence of DNA but the sensitivity is decreased. Poly(A) can be detected in as little as 2 ng of mRNA.
Published Version
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