I'm optimising a protocol for coral mucus sampling and DNA extraction for 16S rRNA gene sequencing. There are many different methods described in papers, mainly by syringes or swabs and different extraction kits being used. Which method minimizes coral tissue contamination and gives the highest DNA yield?
We cannot confirm by molecular analysis, but we collected some mucus using needle-less syringes connected with a short plastic tip to minimize the tissue collection. However, it is very difficult to collect "mucus only" samples, as with sampling you also get seawater at DBL level. You might cite that in the manuscript later.
ciao Walter!
Thanx, yes it is impossible to separate the two. The idea is to sample coral tissue and water column samples too and include those in the sequencing analysis.
Hello Alessandra
The method depends on the coral species you want to sample. In general, you can extract liquid mucus from the Acropora colonies by exposing it in air for some time. In this method, small branches of the colonies will be collected and exposed to air. The mucus secreted for the first five minutes will be washed off with filter-sterilized seawater. The mucus secreted thereafter will be collected by inverting the branches into the sterile tubes. After collection, the branches can be planted back in the reef. This is one of the less destructive ways of sampling. However, this method will not work in the case of boulder-type coral like Porites. These corals secrete mucus as sheets that can be collected underwater using syringes without needles. Contamination with seawater is unavoidable. Therefore you have to collect seawater from the sampling location and sequence it to identify the contaminants
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