The KAPA Stranded RNA-Seq Kit with RiboErase is available for use with human, mouse, and rat samples and is robust and reproducible with flexible input amounts.
Sequences were obtainable from all 18 rat samples (100% success rate) with the RAG1 A primers.
RNA preparation was done using the same purification protocol as for the rat samples above.
Indeed, Leptospira was isolated only from rat samples (R. rattus and R. norvegicus) collected in Toamasina and Toliara.
Even for the 1 year-old specimens, RNA integrity was significantly inferior to the rat samples after 1 year storage at 37°C (Fig. 5a, and Fig. 1).
MPO-DNA complexes were quantified in human and rat samples (conditioned medium and plasma) by combining two different ELISA tests as previously described [31].
If, as alleged by Durham [7], fleas from ship-borne black rats introduced in 1899 were the transmission vector, endemic rat samples collected prior to 1899 should be free of trypanosome infection.
Finally, NETs contributed, at least in part, to cf-DNA measured in plasma and conditioned medium since MPO/DNA complexes were detected in higher amounts in P. gingivalis-infected rat samples (Figure 11B).
Digestion with either exogenously added human caspase-3 or activation of the endogenous salmon caspase with dATP and cytochrome C, resulted in the production of a 110 kDa (SBDP110) not seen in the rat samples.
The rat samples were collected under conditions approved by the Danish Agency for Protection of Experimental Animals and by the inhouse Animal Welfare Committee of the Institute for Food and Veterinary Research, DTU.
Gene-specific primers were the following: KCR 1 for rat samples: forward-primer: 5'-TTC AGG AAG ATA CAG CCC AGA-3', backward-primer: 5'-GGG TTG GAA ATA CTG CTA GGG-3'; KCR1 for pig samples: forward-primer: 5'-GAC GAG ATC TTC CAC CTG C-3', backward-primer: 5'-TAG AAG TTG CCA ACA CTG AAG-3'.
