Question: Low value of mapped reads against a metagenomic co-assembly
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gravatar for Biok
6 months ago by
Biok0
Biok0 wrote:

Hi, everyone, I have two metagenomic samples, I co-assembled these samples together using both spades and Megahit and then I mapped each sample against the co-assembly using bowtie2. I got for each sample around 40 % overall alignment rate. I would like to know if this low value is normal (given that the assembly comes from reads of both samples)???

alignment next-gen assembly • 249 views
ADD COMMENTlink modified 6 months ago by biobiu110 • written 6 months ago by Biok0

By default, megahit have a contig length cut-off of 200 base pairs, so a lot of reads may have been left out of the final assembly.

You can change the minimal contig length with the --min-contig-len command-line option:

 --min-contig-len  <int>  minimum length of contigs to output, default 200
ADD REPLYlink written 6 months ago by h.mon29k

thanks for the reply, I selected contigs larger than 1000 bp because my goal is to reconstruct Metagenome-assembled genomes (MAG's). Initially I got around 7,000,000 contigs from Megahit but after selecting contigs larger than 1000 bps I only got an final assembly of around 455,000 contigs and I used this assembly to map the filtered reads from each sample and I got 40 % overall alignment rate. Many contigs were removed, maybe that's the reason that explains the overall alignment rate.

ADD REPLYlink written 6 months ago by Biok0
0
gravatar for biobiu
6 months ago by
biobiu110
United States
biobiu110 wrote:

Its not necessarily low...

  1. If it's host metagenomes you might have the host reads. Try mapping to the host genome.

  2. If it doesn't help much would try assembly separately to each sample.

  3. Remember that the assembly quality is affected by the read length, sequencing depth and the diversity of the samples.

Good luck :)

ADD COMMENTlink modified 6 months ago • written 6 months ago by biobiu110
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