title: Quantification of miRNA by poly(A)-RT-qPCR arrays and verification of target sites in HIV-1 using a one-LTR infectious molecular clone.
abstract: Quantitative PCR (qPCR) provides a robust method for quantifying DNA species. By combining modern qPCR techniques with the isolation of small RNA, the polyadenylation of the RNA, and the use of reverse transcriptase to create miRNA derived cDNA, it is now possible to use qPCR to quantify miRNA. This method is scalable and provides a useful addition to the retrovirologists' toolbox. Here, we also describe the use of one-LTR infectious molecular clones to verify miRNA target sites within the retroviral LTR.
authors.affiliation: National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Zachary A
authors.initials: ZA
authors.lastname: Klase
authors.forename: Laurent
authors.initials: L
authors.lastname: Houzet
authors.forename: Kuan-Teh
authors.initials: KT
authors.lastname: Jeang
mesh_terms: D003001:Cloning, Molecular; D057809:HEK293 Cells; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics* / Q000302:isolation & purification; D011061:Poly A / Q000235:genetics*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D020079:Terminal Repeat Sequences / Q000235:genetics*
chemical_list: D035683:MicroRNAs; D011061:Poly A