How do you purify a specific RNA?
There are various approaches to RNA purification including phenol-chloroform extraction, spin column purification, and the use of magnetic beads. Total RNA purification involves the extraction and purification of total RNA from your sample, for use in gene expression analyses such as RT-qPCR or RNA-seq.
How is RNA extracted from paraffin embedded tissue?
RNA extraction from paraffin-embedded tissues is made up of four steps. The first one is the deparaffinization and hydration of the tissue sections. The second is the digestion with a proteolytic enzyme such as proteinase K in the presence of effective RNase inhibition to remove proteins from the samples.
How is RNA extracted and purified?
Lysed samples treated with RNase inhibitors and high salt concentrations are passed through the spin columns during centrifugation, while the RNA remains bound to the silica within the column. Washing removes impurities, then the purified RNA is eluted from the silica filter using water.
How do you measure RNA quality?
The most common method used to assess the integrity of total RNA is to run an aliquot of the RNA sample on a denaturing agarose gel stained with ethidium bromide (EtBr). While native (non-denaturing) gels can be used, the results can be difficult to interpret.
How do you store tissue for RNA extraction?
If you do not have or are worried of the affect the RNA stabilizers may have on your sample, you should snap freeze your tissue as soon as possible–either by liquid nitrogen or ethanol on dry ice (LN2 is better). If you give your tissue overnight, the RNA will be worthless for most studies.
Is the rneasy FFPE kit suitable for RT-PCR?
The RNeasy FFPE Kit isolates all RNA molecules longer than 70 nucleotides from FFPE samples, providing usable RNA fragments for numerous downstream applications, including RT-PCR. However, RNA purified from FFPE samples is heavily fragmented and should not be used in downstream applications that require full-length RNA.
Why is lysis buffer used in rneasy FFPE kit?
The lysis buffer efficiently releases RNA from FFPE tissue samples, preventing further RNA degradation. These optimized conditions allow purification of all usable RNA, leading to greater yields from FFPE samples with the RNeasy FFPE Kit than with other methods.
How is the Maxwell RSC RNA FFPE kit used?
The Maxwell® RSC RNA FFPE Kit is used with the Maxwell® RSC Instruments to provide an easy method for efficient, automated purification of RNA from mammalian formalin-fixed, paraffin-embedded (FFPE) tissue samples. The kit does not require the use of hazardous organic solvents, such as xylene, ensuring a safer protocol than other common methods.
How does the PureLink FFPE RNA isolation kit work?
The PureLink FFPE RNA Isolation Kit provides a simple and rapid method for isolating total RNA from formalin-fixed paraffin-embedded (FFPE) samples with high-quality results. A fast and easy heating procedure replaces traditional labor-intensive protocols that use toxic organic solvents for deparaffinization and give disappointing results.