Box 1 | Common statistics for describing genome assemblies

From the following article:

A beginner's guide to eukaryotic genome annotation

Mark Yandell & Daniel Ence

Nature Reviews Genetics 13, 329-342 (May 2012)


Genome assemblies are composed of scaffolds and contigs. Contigs are contiguous consensus sequences that are derived from collections of overlapping reads. Scaffolds are ordered and orientated sets of contigs that are linked to one another by mate pairs of sequencing reads.

Scaffold and contig N50s

By far the most widely used statistics for describing the quality of a genome assembly are its scaffold and contig N50s. A contig N50 is calculated by first ordering every contig by length from longest to shortest. Next, starting from the longest contig, the lengths of each contig are summed, until this running sum equals one-half of the total length of all contigs in the assembly. The contig N50 of the assembly is the length of the shortest contig in this list. The scaffold N50 is calculated in the same fashion but uses scaffolds rather than contigs. The longer the scaffold N50 is, the better the assembly is. However, it is important to keep in mind that a poor assembly that has forced unrelated reads and contigs into scaffolds can have an erroneously large N50. Note too that scaffolds and contigs that comprise only a single read or read pair — often termed 'singletons' — are frequently excluded from these calculations, as are contigs and scaffolds that are shorter than ~800 bp. The procedures used to calculate N50 may therefore vary between genome projects.

Percent gaps

Another important assembly statistic is its percent gaps.Unsequenced regions between mate pairs in contigs and between scaffolds are often represented as runs of 'N's in the final assembly. Thus two assemblies can have identical scaffold N50s but can still differ in their percent gaps: one has very few gaps, and the other is heavily peppered with them. Estimates of gap lengths are often made based on library insert sizes and read lengths; when these are available, the number of 'N's in these gaps usually, but not always, represents the most likely estimate of that gap's size; sometimes, all gaps are simply represented by a run of 50 'N's regardless of their size.

Percent coverage

Percent coverage is used in two senses: genome coverage and gene coverage. The first number, genome coverage, refers to the percentage of the genome that is contained in the assembly based on size estimates; these are usually based on cytological techniques116, 117. Genome coverage of 90–95% is generally considered to be good, as most genomes contain a considerable fraction of repetitive regions that are difficult to sequence. So it is not a cause for concern if the genome coverage of an assembly is a bit less than 100%. Gene coverage is the percentage of the genes in the genome that are contained in the assembly. Gene and genome coverage can differ from one another, as hard-to-assemble repetitive regions are often gene-poor. As a result, the percentage gene coverage is often substantially larger than the percentage genome coverage for some difficult-to-assemble genomes.