Astrangia Nutrition DNA Extractions
Using the Zymo Quick-DNA Mini Prep kit on adult Astrangia poculata samples from the Astrangia Nutrition Experiment.
Experimental design
All samples were snap frozen on 20190417 and stored at -80C. 8 randomly selected corals were airbrushed on 20190418 and aliquoted appropriately. DNA extractions were performed on the same day.
Table 1: A. poculata homogenate samples used for DNA extractions.
Vial # | Coral ID | Volume of Sample (mL) |
---|---|---|
13 | 2753 | 0.5 |
19 | 1709 | 0.5 |
25 | 1811 | 0.5 |
31 | 1275 | 0.5 |
37 | 1756 | 0.5 |
43 | 2014 | 0.5 |
49 | 2189 | 0.5 |
55 | 2564 | 0.5 |
Methods
- All samples were stored at -20C until all corals were airbrushed and ready to be extracted for DNA.
- Once thawed, all samples were centrifuged to separate the host and symbionts.
- 200uL of the supernatant (host) was transferred to a new 1.5mL tube and used for the DNA extraction.
- The rest of the extraction followed this protocol
Quantification
Quantified DNA concentration using 1 uL of sample on the Qubit
Table 2. Qubit quantification
Standard 1 | Standard 2 | #13 (ng/uL) | #19 (ng/uL) | #25 (ng/uL) | #31 (ng/uL) | #37 (ng/uL) | #43 (ng/uL) | #49 (ng/uL) | #55 (ng/uL) | |
---|---|---|---|---|---|---|---|---|---|---|
DNA | 209.29 | 22686.75 | 31.6 | 9.28 | 31.0 | 43.4 | 44.2 | 27.6 | 29.0 | 19.3 |
DNA | - | - | 31.6 | 9.30 | 31.0 | 43.2 | 44.6 | 27.6 | 28.8 | 19.3 |
Written on April 18, 2019