Montipora Larvae DNA/RNA Extraction Batch 2

Larvae from Biomineralization Project


For detailed extraction protocol view post: Protocol for DNA/RNA Extractions of Montipora Coral Larvae Using Zymo-Duet Extraction Kit


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 TS-batch-2-3-sample-info.png TS-biomin-Ext-Batch-2-14-15.png TS-biomin-Ext-Batch-2-16-17.png

Rerun of sample 363:

TS-biomin-Ext-Batch-2-16.png)

Gel

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

annotated-biomin-gel-batches-2-3.png

Written on March 21, 2019