How can I avoid or remove genomic DNA contamination from the total RNA preparation?
Carry out all procedures in a “DNA-free” workspace (see the first Sample Preparation FAQ, above). Be sure to include any DNase treatment steps in the recommended RNA isolation procedure or treat RNA separately with RNase-free DNase followed by re-purification using a spin-column based method. Be sure to double both the units of enzyme and the incubation time recommended by the RNase-free DNase manufacturer. When isolating RNA from tissue samples, it is recommended that the tissue be subject to Trizol extraction (Invitrogen) prior to DNase treatment and RNA isolation. In order to minimize DNA contamination in your RNA preparations, and avoid the need for supplemental DNase treatments, we recommend using the SABiosciences RT2 First Strand Kit (C-03), which includes a highly efficient genomic DNA elimination step before reverse transcription.
Related Questions
- Which costs, especially with regards to reagents, are to be expected for the complete experiment (from total genomic DNA isolation till pathogen detection)?
- Does the total genomic DNA need pre-treatment before its application on the LOOXSTER® universal columns?
- How can I avoid or remove genomic DNA contamination from the total RNA preparation?
