Compositions and methods for identifying and comparing members of microbial communities using amplicon sequences
In alternative embodiments, the invention provides computational algorithms, computer programs, software and other methods, systems and products of manufacture (e.g., computers, devices or apparatus) for identifying members of microbial communities, their abundance and distribution from amplicon sequence data, and comparing microbial communities and consortia. In alternative embodiments, the invention provides computer-implemented methods comprising a subset of, substantially all, or all of the steps as set forth in the flow chart of FIG. 1 , FIG. 3 or FIG. 4 . In alternative embodiments, the invention provides methods for identification of consortia, optionally followed by construction of artificial microbial consortia from pure strains or enrichment cultures. In alternative embodiments, the invention provides compositions, fluids, bioreactors, muds, reservoirs or products of manufacture comprising a synthetic microbial consortium made by the method of the invention.
1. A method for making a synthetic microbial consortium comprising:
a) providing a plurality of nucleic acids from one or more samples;
b) sequencing the nucleic acids;
c) processing the sequencing reads into tags by trimming the sequencing reads at a defined length, wherein sequencing reads shorter than the defined length are discarded;
d) quality filtering the tags by removing ambiguous truncated sequencing reads or sequencing reads that falls below a quality threshold;
e) determining the count abundances of tags in the one or more samples;
f) importing the sequences and identifiers of new tags into a database;
g) importing the count abundances of tags in samples into the database;
h) exporting data of the tag count abundance of at least two samples from the database, wherein a threshold for minimum tag abundance is set for tags to appear in a correlation analysis;
i) creating a data output comprising description of microbial communities identified as counts of abundance of unique members of each community; and
j) combining microbial cultures corresponding to the composition of microbes correlated with environmental distributions to create a synthetic microbial consortium.
2. A synthetic consortium made by the method of claim 1 .
3. A composition, a fluid, a bioreactor, a mud, a reservoir, or a product of manufacture comprising a synthetic microbial consortium made by the method of claim 1 .
4. The method of claim 1 , wherein the one or more samples comprise bacteria, archaea, eukaryotes, or viruses.
5. The method of claim 1 , wherein the sequencing comprises amplification of the nucleic acids, wherein the amplification uses primers comprising a barcode.
6. The method of claim 5 , wherein the barcode corresponds to a sample or an environmental parameter.
7. The method of claim 1 , wherein the sequencing associates each base with a quality score.
8. The method of claim 1 , wherein the nucleic acids from the one or more samples are sequenced simultaneously.
9. The method of claim 1 , wherein the nucleic acids from the one or more samples are sequenced on multiple instrument runs.
10. The method of claim 1 , wherein the trimming of the sequences comprises recognizing conserved sequence patterns within the sequencing read.
11. The method of claim 1 , wherein the tags are taxonomically classified.
12. The method of claim 1 , wherein the tag abundances represent absolute counts or a fraction of total sequences obtained from a sample.
13. The method of claim 1 , wherein the abundance data is log transformed.
14. The method of claim 1 , wherein the data is exported in a human-readable format.
15. The method of claim 1 , further comprising identifying the composition of microbes correlated with environmental factors, wherein each tag is representative of a particular group of microbes with correlated environmental distributions.
16. The method of claim 1 , further comprising constricting the database.