Reverse transcription-polymerase chain reaction (RT-PCR) is a process that transforms RNA to cDNA via a reverse transcriptase, then replicates it with a DNA polymerase. To get good results from reverse transcription, the starting RNA material must be of sufficient quantity, intact, and free of genomic DNA.

Real-time PCR—also known as quantitative PCR or qPCR—quantifies the copy number of PCR templates, such as DNA or complementary DNA (cDNA). Invitrogen kits and reagents for RNA extraction for qPCR are designed to yield the quality and quantity of RNA you need for RT-PCR reactions

Learn the steps involved in RNA isolation

Selection guide: RNA extraction kits for RT-PCR and qPCR

 Gold standard for highly pure, intact RNAHigh-quality RNA in less than 20 minutesHigh-throughput purification of RNA and DNAComplete, no purification system for qRT-PCR results
Type of PCR supportedReverse transcription PCR and downstream applicationsReverse transcription PCR and downstream applicationsReverse transcription PCR and downstream applicationsStraight to qPCR
Reagent/KitTRIzol reagentsPureLink kitsMagMAX kitsCells-to-CT kits
BenefitsMost cited, economicalQuick and convenientScalable and flexibleStreamlined, best for qRT-PCR
Isolation methodOrganic reagentSilica filter, column or plate formatMagnetic beads (scalability, flexibility)Chemical lysis
High throughput compatibleNoNoYesYes
RT reagents includedNoNoNoYes
qPCR reagents includedNoNoNoYes
Prep time ~1hr< 20 minutes~45 minutes10 minutes
Compatible sample typesMost sample types, including difficult to lyseCells and tissuesCells, blood, plants, tissueCells
Amount of starting material100 mg of tissue or 107 cells (requires 1 mL reagent)Up to 200 mg tissue or 5 x 106 to 1 x 108 cellsUp to 100 mg tissue or up to 5 x 106 cells1–103 cells

 

Purifying RNA samples prior to qPCR: DNA removal

RNA samples should be purified prior to qPCR. Genomic DNA contamination can lead to false positive RT-PCR results. Thermo Fisher Scientific offers a number of DNases that can be used to eliminate genomic DNA that may be present in your RNA preparation.

The DNA-free DNase Treatment and Removal Kit contains everything you need to remove contaminating DNA from RNA samples and remove the DNase without the use of Proteinase K treatment and organic extraction.

The TURBO DNase enzyme kit includes a hyperactive enzyme engineered from wild-type bovine DNase. The proficiency of TURBO DNase enzyme in binding very low concentrations of DNA means that the enzyme is particularly effective in removing trace quantities of DNA contamination.

Resources

Stylesheet for Classic Wide Template adjustments