Total RNA was purified from 10-fold serial dilutions of Jurkat cells (5 x 105 – 5 cells) using the RNeasy UCP Micro Kit. The β-actin transcript was detected inas few as 5 cells by real-time RT-PCR. In control reactions without reverse transcriptase (-RT), β-actin DNA
was not detected after 40 cycles, indicating the absence of genomic DNA
Sample preparation was performed with mock samples to check for possible nucleic acid contamination in the kit. Three independent experiments with mock preparations were done with 3 different lot numbers for the RNeasy Micro Kit, as well
as for the RNeasy UCP Micro Kit. Eluates were analyzed by RT-PCR using primers for a
conserved bacterial 16S rRNA sequence. Delta CT values between the no template contro (NTC) and the empty
preparation were calculated for both kits. The delta CT value for the RNeasy UCP Micro Kit was lower compared to the delta CT of the RNeasy Micro Kit, showing that there are
minimal contaminants in the RNeasy UCP Micro Kit.
Total RNA was purified from 10‐fold serial dilutions of Jurkat cells
using the RNeasy UCP Micro Kit. The β‐actin transcript was detected even down to one cell by real‐time
RT‐PCR, and reproducible CT values were observed at each dilution, indicating the absence of inhibition.