App Note
A rapid, single-tube protocol for extracting RNA from cell culture
Most methods for extracting RNA from cell culture or small tissue samples rely on solid phase purification or solvent-based extraction and precipitation. In some cases, lysis is achieved using agents that must be neutralised or removed from the solution before the nucleic acid is usable for analysis. This requirement increases the number of manipulations; and if solid phase or solvent methods are used, yields are reduced and bias can be introduced into the population of mRNAs.
RNAGEM uses a rapid, single-step protocol that releases RNA and DNA with excellent linearity across a wide range of cell numbers, from a single cell to thousands of cells. The method is automatable, closed-tube and does not require further purification of the RNA for accurate RT-qPCR analysis. The RNAGEM reagents efficiently lyse the cells and strip protein complexes from nucleic acids, thereby allowing higher processivity of polymerases. The result is greater sensitivity – especially with low abundance transcripts. Reduced handling, and efficient template preparation means that the RNAGEM kits generate mRNA profiles that are as close to the biological reality of the sample as possible.
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RNAGEM
Temperature-driven, single-tube extractions of total RNA from mammalian cells, tissues, insects, bacteria and virus
This powerful, broad-specificity approach is ideal for gene expression from a very small number of cells, such as exosomes or single cells, without the need for harsh chemicals or further purification.
- Ideal for preparing RNA from mammalian cell culture, laser capture microdissection, FACS-prepared cell populations, cells in suspension, adherent cells, cells stored in RNAlaterTM and cell pellets
- Simplified, hands-off workflow provides RT-PCR and RT-qPCR ready RNA in approximately 5 minutes
- Releases both RNA and DNA with excellent recovery across a wide range of cell numbers
- Requires no further purification for accurate RT-PCR and RT-qPCR analysis
- Inhibitor-free means no harsh chemical washes or multiple steps
- RT-PCR and RT-qPCR reagents can be added directly to the lysate for a simple, streamlined protocol in 96 well plates
- Each kit contains RNAGEM, DNase I, BLUE buffer, 10x DNase buffer, and 10x TE
Learn more about RNAGEM and recent applications. Download our RNAGEM technical overview. Watch our webinar on advances in extracting and stabilizing RNA.