Figure 2. Good Amplification and Consistency of Isolation. Norgen's Total RNA Purification Maxi Kit was used to isolate total RNA from six different overnight samples of E. coli (10 mL samples). In brief, one microgram of the isolated RNA was subjected to a 20 µL reverse transcription using Invitrogen Superscript III. Three microliters of the reverse transcription were used as the template in a 20 µL qPCR reaction using 16S rRNA E. coli specific primers and Bio-Rad iQ mix. As it can be seen, the RNA was detected and successfully amplified in all cases. Furthermore, all samples showed a similar Ct value, indicating consistent isolation.