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 3, 2019

Samples

Extraction #	Sample #	Date Collected	Quantity (µL)	Extracter
14	244	6/17/2018	300	Erin
15	243	6/17/2018	300	Erin
16	363	6/22/2018	300	Erin
17	310	6/18/2018	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 15 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.
• The cap broke on RNA elute 310. Tranferred to new 1.5 mL centrifuge tube.

Qubit
Quantified on March 21, 2019 with Batch 3

• 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
14	244	192.3	19059	27.8	27.2	27.5	406.9	10912	110.0	109.0	109.5
15	243	192.3	19059	41.8	40.4	41.1	406.9	10912	105.0	105.0	105.0
16	363	192.3	19059	38.2	38.4	38.3	406.9	10912	244.0	242.0	243.0
17	310	192.3	19059	67.0	65.2	66.1	406.9	10912	85.4	86.0	85.7

Tape Station

• Followed RNA TapeStation protocol
• Low RIN for sample 363 because not add enough sample to well H1. Reran on April 1, 2019

Rerun of sample 363:

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Gel

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