I need to perform metagenomics on flocs containing bacteria, fungus and microalage, but I only have a software that can do etagenomics analyses for 16s bacteria.
Hi, do you have rRNA amplicon data or total DNA metagenomic data ? For the first you can use Mothur http://www.mothur.org/wiki/Main_Page, it can deal with 16S and 18S rRNA data. For the second case, you can still use Mothur for sequence processing, and for data analysis MGRAST online http://metagenomics.anl.gov/ or MEGAN http://www-ab.informatik.uni-tuebingen.de/software/megan5 on your own computer.
For the fungal metagenoimc analysis (Phylogenetic/taxonomic analysis), you can use internal transcribed spacer (ITS) for the amplification/sequencing in place of 16S rdna (for bacterial).
1. Analysis of reads can be done using RDP classifier but you have to use separate database for fungal 18S/ITS sequences for example FungiDB.
2. Analysis of the reads can be done using MEGAN software.
http://ab.inf.uni-tuebingen.de/software/megan5/
QIIME can be used for the OTU analysis. You can refer the mentioned link below http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3712477/pdf/nph0199-0288.pdf
You can also use CloVR-ITS see the paper mentioned below.
CloVR-ITS: Automated internal transcribed spacer amplicon sequence analysis pipeline for the characterization of fungal microbiota
Regarding the shotgun metagenomic sequence analysis you can use MG-RAST pipeline.
As Fabian already said, you could make use of the MG-Rast software for metagenomics datasets. It is really friendly in use and had a clear overview manual. To upload the data everything is also explained step by step. I found MG-Rast a good tool to start with.
If you have an amplicon dataset and it's not to large, you could also make use of SILVA-NGS. It doesn't exist that long, but gives you some images about the taxonomy where you could start with.
Several of these softwares are available free on the net like MEGAN, MOTHUR etc. They all come with detailed descriptions as to the purpose they are to be used for. Check and see which is most suitable for you. Good luck.
According to MG-RAST manual, it is designed for prokaryotic metagenomics analysis "The system supports the analysis of the prokaryotic content of samples, analysis of viruses and
eukaryotic sequences is not currently supported, due to software limitations".