High-quality RNA.

Concentration of RNA.

Reliable cleanup of RNA.

Efficient removal of real-time RT-PCR inhibitors.

Efficient removal of real-time RT-PCR inhibitors.

Total RNA was isolated from HeLa cells using an acid-phenol method plus cleanup and concentration using the RNeasy MinElute Cleanup Kit. The high quality of the RNA is shown by scanning with the Agilent 2100 Bioanalyzer.

The indicated amounts of total RNA from HeLa cells were concentrated using the RNeasy MinElute Cleanup Kit, and 5 µl of the 12 µl eluate was used for each reaction. Real-time RT-PCR was carried out on an ABI Sequence Detection System using the QuantiTect Probe RT-PCR Kit, with primers and probes specific for β-actin.

The indicated amounts of total RNA from HeLa cells were concentrated using the RNeasy MinElute Cleanup Kit. Real-time RT-PCR was carried out on the LightCycler System using the QuantiTect Probe RT-PCR Kit, with primers and probes specific for the p16 gene.

Efficient removal of real-time RT-PCR inhibitors.

Without cleanup, inhibitors can lead to underestimation of RNA amounts in real-time quantification. Total RNA was isolated from HeLa cells using an acid-phenol method, with or without subsequent cleanup using the RNeasy MinElute Cleanup Kit. Real-time RT-PCR was carried out as described in figure "Concentration of RNA", with the indicated volumes of the RNA eluate. Apparent RNA amounts were calculated from the resulting C_T values.