Eggs and sperm from FACE PUF Morea 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: May 17, 2019

Samples

ID	Spawning Date	Content	Sample Quantity (µL)	Sample Conserv.	Tech
120	2/11/2018	Eggs	100	300 uL Shield	Erin
797	4/11/2018	Eggs	100	300 uL Shield	Erin

Extraction Notes

• Samples were already preserved in 300µL DNA/RNA Sheild so did not add more
• Digested for 3 hours and 40 minutes at 55°C at speed 35 in Incubator-Genie because test tubes were too large for the thermomixer
• Added additional 15µL Proteinase K after 2 hours and 50 min
• Some lipids left in samples
• Pelleted at 2300 rcf for 1 min

Qubit
Quantified May 22, 2019

• 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
73	120	184.2	20357	Too Low	Too Low	Too Low	385.6	10602	60.4	60.4	60.4
74	797	184.2	20357	Too Low	Too Low	Too Low	385.6	10602	88.2	88.0	88.1

Tape Station

• Followed RNA TapeStation protocol