Molecular biology methods are highly sensitive to detect the genome of pathogens and to study their biology. Polymerase chain reaction (PCR) and reverse transcription followed by a polymerase chain reaction (RT-PCR) permit the detection of the presence and the replication of African swine fever virus in soft ticks. Here, we described our techniques to detect and quantify DNA and RNA of African swine fever virus in soft ticks including a housekeeping gene of soft ticks as internal control.