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: April 29, 2019

Samples

Extraction #	Sample #	Date Collected	Quantity (µL)	Extracter
45	133	6/13/2018	300	Erin
46	153	6/13/2018	300	Erin
47	155	6/13/2018	300	Erin
48	158	6/13/2018	300	Erin
49	160	6/13/2018	300	Erin
50	166	6/14/2018	300	Erin
51	169	6/14/2018	300	Erin

Extraction Notes

• Gathered all samples and set aside on ice. Then added DNA/RNA Shield to all.
• Digested for 2 hours and 12 minutes at 55°C
• All samples except for 166 had very high lipids.
• After digestion, pelleted out lipids in all samples. Centrifuged at 2200 rcf for 1 min.
• Dropped sample #169 after adding EtOH (Step 2 RNA Extraction). Spilled about 1/2 of sample.

Qubit
Quantified on May 10, 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
45	133	143.3	18426	Too Low	Too Low	Too Low	389.4	11715	80.0	79.8	79.9
46	153	143.3	18426	Too Low	Too Low	Too Low	389.4	11715	47.2	47.2	47.2
47	155	143.3	18426	2.16	2.10	2.13	389.4	11715	216	216	216
48	158	143.3	18426	Too Low	Too Low	Too Low	389.4	11715	129	129	129
49	160	143.3	18426	Too Low	Too Low	Too Low	389.4	11715	45.2	45.6	45.4
50	166	143.3	18426	Too Low	Too Low	Too Low	389.4	11715	79.0	79.0	79.0
51	169	143.3	18426	4.64	4.62	4.63	389.4	11715	34.8	34.4	34.6

Tape Station

• Followed RNA TapeStation protocol

Tape stationed May 22, 2019