Assemblathon, GAGE and Four Dimension of Evaluating Genome Assembly Programs

Assemblathon, GAGE and Four Dimension of Evaluating Genome Assembly Programs

We wrote about the Assemblathon paper in January, when it was first uploaded in arxiv (a place declared as ‘low grade journal’ by omniscient Andrew Gilman).

Notes on Assemblathon Paper

Titus Browns Thoughts on the Assemblathon 2 paper

Based on tweets from @assemblathon, the paper will be out in ‘print’ soon.



Speaking of comparison between assembly programs, our reader Anton Korobeynikov emailed us a commentary that the readers will find informative.

The recently published GAGE-B paper (Magoc, et al., 2013) presents an evaluation of several popular assemblers, including SPAdes 2.3.

Since SPAdes 2.5 is out, we evaluated it on the data sets from the GAGE-B study. Four MiSeq data sets (B. cereus, R. sphaeroides, M. abscessus, and V. cholerae) were selected for the assessment. Since these reads are 250 bp length, we applied our recommendations for assembling long Illumina paired-end reads with SPAdes.

The original coverage of those data sets is about 500x. However, in the GAGE-B experiments, all data was down-sampled to 100x coverage, because higher coverage barely affected contig size. Meanwhile, SPAdes benefits from high coverage, so we decided to assemble the data sets with the original ~500x coverage. Our tables contain the GAGE-B assemblies of 100x-coverage data, and the SPAdes 2.5 assembly of 500x-coverage data.

The B. cereus data was downloaded from the official Illumina website. The other three data sets were obtained from the Sequence Read Archive at NIHs National Center for Biotechnology Information (NCBI): SRR522246 (R. sphaeroides), SRR768269 (M. abscessus), SRR769320 (V. cholerae). Genome references and contigs produced by other assemblers mentioned in the GAGE-B study were downloaded from the GAGE-B website.

It includes a table of comparison between various genome assembly algorithms done in the GAGE-B style. The numbers for SPADES improved a lot compared to what was seen in GAGE-B paper.

SPAdes and MaSuRCA Assemblers Performed Best in GAGE-B Evaluation

Those seeking information on Ray and few other assemblers not included in the above GAGE-B related link should check here.


That brings us to the question of what Assemblathon-3 (or GAGE-C) should be like. The comparisons are very informative for the community, but it would be great, if three issues are addressed.

Four Dimensions of Evaluating Genome Assembly Programs

Users of bioinformatics programs care about three issues -

(i) Quality of results,

(ii) Time taken to get the answer,

(iii) Cost of analysis.

We split time and cost as separate metrics, because it is not possible to run some assembly programs without 256 Gb or 512 Gb RAM, no matter how long you are willing to wait. Assemblathon focused on the quality issue, but the other two factors are quite important.

iv) Fourth dimension - polymorphic genomes

In trying to understand the genome assembly programs, we used to focus on those three dimensions (quality, time, cost), until we came across the pacific oyster genome. You can see how different the oyster genome is from others in the preqc charts of Jared Simpson.

Very Helpful Preprocessing Module for Those Interested in Assembling Genomes

Even excluding the other complexities such as metagenome, transcriptome or single cell assembly, a comparison of assemblers need to address those four dimensions to be fully informative.

Rapidly changing nature of field

Between the time the Assemblathon entries were submitted and the paper will be formally published, many assembly algorithms changed a lot. To be useful to the community, evaluations will possibly have to publish a lot in arxiv and leave the ‘high-grade’ journals to Andrew Gelmans. In that way, SPAdes blog has been very helpful, because they continually update their benchmarks with the latest versions of other assemblers. For alignment programs, nothing is as informative as the often updated ROC curves from Heng Li’s website. The BWA-MEM paper also includes information on run time for the programs.

Written by M. //