title: Single-cell cDNA-PCR: removal of contaminating genomic DNA from total RNA using immobilized DNase I.
abstract: A procedure utilizing immobilized DNase I that allows the efficient amplification of cDNA by PCR from a single cell in the absence of contaminating genomic DNA is described. DNase I treated, total RNA derived from single cells was reverse transcribed into cDNA followed by PCR using beta-actin and c-fos specific primers that recognize different exons of the respective genes. Amplification products corresponding to cDNA, but not to genomic sequences, were detected after treatment with immobilized DNase I in samples previously shown to be contaminated with genomic DNA. This method allows the efficient removal of DNA contaminating total RNA derived from a single cell.
authors.affiliation: Dept. of Clinical Physiology & Occupational Medicine, University of Ulm, FRG.
authors.forename: B L
authors.initials: BL
authors.lastname: Ziegler
authors.forename: C
authors.initials: C
authors.lastname: Lamping
authors.forename: S
authors.initials: S
authors.lastname: Thoma
authors.forename: C A
authors.initials: CA
authors.lastname: Thomas
mesh_terms: D016477:Artifacts; D001483:Base Sequence; D004247:DNA / Q000032:analysis*; D003850:Deoxyribonuclease I; D004800:Enzymes, Immobilized; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*
chemical_list: D004800:Enzymes, Immobilized; D012333:RNA, Messenger; D004247:DNA; D003850:Deoxyribonuclease I