Goal

To test ITS2 sequencing prep on Porites astreoides and Montipora capitata samples from the Porites July Bleaching, Symbiont Integration, and Bleaching Pairs projects. The DNA from these samples were extracted with the Zymo Quick-DNA kit and Zymo DNA/RNA mini prep kit plus.

The dilution and plate planning can be found here

Protocol

I am using a modified version of this protocol written by E. Strand.

The main modifications from this protocol were to adjust the dilutions for 2 replicates instead of 3.

The negative control was ultrapure water and the positive control was the same positive control from this ITS2 run (Becker E9) diluted to 2ng/ul.

Master Mix Calculation

Plate and Strip Tube layout

Gel Results

Gel Electrophoresis