Larvae from Biomineralization Project

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For detailed extraction protocol view post: Protocol for DNA/RNA Extractions of Montipora Coral Larvae Using Zymo-Duet Extraction Kit

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Extraction Date: March 29, 2019

Samples

Extraction #	Sample #	Date Collected	Quantity (µL)	Extracter
26	311	6/18/2018	300	Erin
27	375	6/22/2018		Erin
28	359	6/22/2018		Erin
29	323	6/20/2018	300	Erin
30	242	6/17/2018	300	Erin

Extraction Notes

• Gathered all samples and set aside in cooler filled with dry ice. Then added DNA/RNA Shield to all.
• Digested for 3 hours and 25 minutes at 55°C
• Added 15 additional microliters of Protienase K after 2 hours and 30 minutes of digestion at 55°C
• After digestion, pelleted out additional tissue in all samples. Centrifuged at 1600 rcf for 1 min.

Qubit
Quantified on April 1, 2019 with Batch 5

• Followed Broad Range dsDNA and RNA Qubit protocol
• All samples were read twice

Extraction #	Sample	DNA Standard 1 (RFU)	DNA Standard 2 (RFU)	DNA 1 (ng/µl)	DNA 2 (ng/µl)	Average DNA	RNA Standard 1 (RFU)	RNA Standard 2 (RFU)	RNA 1 (ng/µl)	RNA 2 (ng/ul)	Average RNA
26	311	179.8	21758	29.2	28.6	28.9	399.8	11414	129.0	129.0	129.0
27	375	179.8	21758	33.0	32.4	33.7	399.8	11414	53.0	52.0	52.5
28	359	179.8	21758	50.2	49.4	49.8	399.8	11414	131.0	132.0	131.5
29	323	179.8	21758	36.4	36.0	36.2	399.8	11414	91.6	92.2	91.9
30	242	179.8	21758	45.6	45.2	45.4	399.8	11414	145.0	144.0	144.5

Tape Station

• Followed RNA TapeStation protocol

Gel

• Followed PPP Lab Agarose Gel protocol
• Used the Thermo Scientific GeneRuler 1 kb Plus DNA Ladder (link)