Extracting DNA/RNA from Cv sperm in seawater

[yaaminiv]

I want to use the Zymo Microprep kit to extract DNA and RNA simultaneously from C. virgincia sperm samples. The sperm samples are in seawater. Based on the kit manual, I assume I'd pellet the sperm cells, remove the supernatant, and proceed with adding the DNA/RNA Lysis Buffer.

@kubu4 or @grace-ac, anything to add based on your experience with the crab hemolymph samples beyond this lab notebook post? I assume that extracting RNA from the crab samples in hemolymph may be similar to trying to extract DNA/RNA from these sperm cells in seawater!

[kubu4]

Are the samples frozen? Any sort of preservative?

My initial concern is that the cells were not pelleted before freezing. Freezing generally lyses cells, which will greatly reduce your yield (if you decide to pellet the cells prior to DNA extraction).

With that being said, what is the sample volume? Are we talking 1mL of sea water or like 100uL? If the volume is small enough, you can process using the Zymo kit (see the manual for info on handling cells in liquid).

If the volume is too large, I'd do an extraction using DNAzol instead.

[yaaminiv]

With that being said, what is the sample volume? Are we talking 1mL of sea water or like 100uL?

Sperm cells were shipped in 100 µL of seawater (no preservative). To clarify, I could follow this protocol?

[kubu4]

That's what I would do.