Real time RT PCR

DNA contamination

Contamination of RNA samples with trace amounts of genomic DNA can interfere with real-time RT-PCR quantification if the PCR primers used are also able to amplify genomic DNA sequences. To avoid the negative effects of genomic DNA contamination, careful primer design is required (see Primer design). If this is not possible, RNA samples should be treated with DNase I to digest contaminating DNA.
Use of appropriate controls will enable the detection of any contaminating DNA in the RT-PCR. Reactions should be set up with and without the reverse transcriptase. The presence of a product in the absence of the reverse transcriptase indicates contamination.
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