Differentiating vector-derived mRNA from contaminating DNA templates by inverse RT-PCR.
Inverse RT-PCR was used for detecting doxycycline-induced mRNA expression after viral transduction with a retroviral vector harboring human TDAG51. After the circularization of double-stranded cDNA, induced transcripts originating from integrated vector genomes were selectively amplified, even in the presence of DNA templates. Thus, DNase treatment before amplification was unnecessary.
Flindt
S
S
Paul Ehrlich Institute, Langen, Germany.
Meier-Noorden
M
M
Hinz
T
T
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004318:Doxycycline / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D019169:Jurkat Cells; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000900:Anti-Bacterial Agents; D012333:RNA, Messenger; D004318:Doxycycline