reversetranscribed

Reversetranscribed is the past participle form used to describe RNA that has been converted into complementary DNA (cDNA) by the enzyme reverse transcriptase. The process, reverse transcription, is a foundational technique in molecular biology and is widely applied in gene expression analysis, cloning, and sequencing workflows. In typical protocols, RNA is isolated from cells and primed with oligo-dT, random hexamers, or gene-specific primers before incubation with reverse transcriptase to synthesize cDNA. The resulting cDNA can then be amplified by PCR, used in quantitative PCR, or prepared for high-throughput sequencing. The term may appear as reversetranscribed RNA or as “RNA was reversetranscribed” in experimental methods.

Reverse transcription is mediated by reverse transcriptase, an RNA-dependent DNA polymerase that synthesizes the first DNA strand complementary to the RNA template. In standard methods, RNase H activity helps remove the RNA strand, allowing second-strand synthesis to create double-stranded cDNA suitable for downstream applications. In retroviruses and retrotransposons, reverse transcription occurs as part of genome replication, with variation and error-proneness affecting sequence accuracy.

Applications of reversetranscribed cDNA include reverse transcription PCR (RT-PCR), quantitative RT-PCR (RT-qPCR), RNA sequencing library preparation, and cloning of transcripts. Limitations include primer- and enzyme-related biases, incomplete conversion of RNA species, and potential artifacts introduced during cDNA synthesis.

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