DNA from Mhf, CMhm and CMt was used as positive controls for the PCR methods . The samples were used as positive controls in the initial analysis to confirm that the used PCR and RT-PCR conditions could produce the expected PCR products. After confirmation, the 16S rRNA PCR fragments from the first haemoplasma-positive blood samples (Mhf and CMhm) were used and cloned using the TOPO TA Cloning® Kit as described by the manufacturer (Invitrogen, Denmark). Since we found no CMt positive cats, we also cloned the CMt 16S rRNA. All cloned fragments were sequenced, confirming that the DNA was of the expected sequence. Plasmid purification was done using the QIAprep® spin miniprep kit from Qiagen. Sequencing of the cloned samples was done using 0.8 µM of each of the primers M13-forward: 5′ GTAAAACGACGGCCAGT 3′ and M13-reverse: 5′ AACAGCTATGACCATG 3′. These plasmids were used as positive controls in the PCR and RT-PCR analysis of the blood samples using the M-forward and M-reverse primers as described under the two PCR methods.