Larvae from Biomineralization Project

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For detailed extraction protocol view post: Testing Number of Montipora Larvae Needed for DNA/RNA Extraction

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Extraction Date: March 22, 2019

Samples

Extraction #	Sample #	Date Collected	Quantity (µL)	Extracter
22	312	6/18/2018	300	Erin
23	231	6/16/2018	300	Erin
24	379	6/22/2018		Erin
25	371	6/22/2018		Erin

Extraction Notes

• Gathered all samples and set aside in cooler filled with dry ice. Then added DNA/RNA Shield to all.
• Sample 231 had larvae frozen to the cap of the test tube. They were centrifuged down after the shield was added.
• Digested for 2 hours and 20 minutes at 55°C
• After digestion, pelleted out additional tissue in samples 371 and 379. Centrifuged at 1600 rcf for 2 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
22	312	179.8	21758	43.0	43.4	43.2	399.8	11414	81.2	80.8	81.0
23	231	179.8	21758	36.8	37.0	36.9	399.8	11414	83.2	82.6	82.9
24	379	179.8	21758	14.1	13.8	13.95	399.8	11414	84.0	83.8	83.9
25	371	179.8	21758	27.8	27.2	27.5	399.8	114142	59.2	59.6	59.4

Tape Station

• Followed RNA TapeStation protocol

Gel

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