Virome shotgun sequencing only reveals the double-stranded DNA (dsDNA) content of a given sample, unless specific treatments are applied. However, genomes of viruses often consist of a circular single-stranded DNA (ssDNA) molecule. Pre-treatment and amplification of DNA using the multiple displacement amplification (MDA) method enables conversion of ssDNA to dsDNA, but this process can lead to over-representation of these circular ssDNA genomes. A more recent alternative employing the xGen kit permits ligation of adaptors directly to sheared and denatured DNA. However, the sonication step might shear ssDNA more efficiently than dsDNA, therefore introducing another bias in virome sequencing. These limitations prompted us to explore an alternative method of DNA preparation for sequencing mixed ssDNA and dsDNA viromes. We present here a new method for sequencing both ssDNA and dsDNA, using the T7 DNA polymerase (T7pol) to convert ssDNA into dsDNA. We compared this method to two others: sMDA with a 30-minute incubation and direct DNA shearing without ssDNA to dsDNA conversion, using a mixture of five bacteriophages, including two with ssDNA. To ensure fairness, all samples were subsequently prepared with the xGen kit. Our fi