Table 2. Properties of Uniquorn compared to established methods for identification of CCLs.

Identification Method for NGS CCLs	Physical sample required	Additional experiments required	Locus coverage required	Zygosity-pattern required	Dependent on reference genome
STR (19)	X	X	-	-	-
SPIA (6)	X	X	X	X	-
NGS SNP (4)	-	-	X	X	X
NGS All Variants (Uniquorn)	-	-	-	-	X

SPIA and STR require additional verification experiments on the physical CCL sample. Identification of CCLs by matching their SNP-zygosities directly from the NGS-data requires that the loci of the characteristic SNPs were sequenced and not filtered. For SPIA and NGS-SNP, zygosity calls have to be comparable (technology, ploidy, algorithms, etc.). Uniquorn only requires using the same reference genome for variation calling. Note that CCL samples created with a specific reference genome versions can be converted into another version, e.g. by a lift-over software, thereby decreasing the severity of this limitation.