Fig. 3

Amplification plots from RT-qPCR analysis on total RNA from fecal samples for glyceraldehyde 3-phosphate dehydrogenase (GAPDH), ribosomal protein S9 (RPS9), toll-like receptor 4 (TLR4). The reverse transcriptase Moloney murine leukemia virus (MMLV) used in this test was SuperScript IV (Cat# 18090050; Invitrogen, Carlsbad, CA, USA), while the genetically modified MMLV was RevertAid (Cat# EP0442; Thermo Scientific, MA, USA). Three fecal samples from different calves were used to isolate total RNA, which was, in turn, used to synthesize cDNA using equal amounts of either reverse transcriptase (i.e., MMLV and genetically modified MMLV). Dashed line in red indicates the threshold with the actual value in red for each gene. Arrow denotes the 30th amplification cycle. Additional protocol information can be found in the Suppl. Materials