Project: Thermal Transplant Molecular

Goal

ITS2 sequencing prep on Porites astreoides adult and larval samples. 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 and each well had a total of 25 uL.

Master Mix Calculation

Dilution Plate planning and PCR Plate Layout

Gel Results

Gel Electrophoresis

• I ran a large 100 well 2% agarose gel of the above protocol and ran it at 100V for 60 minutes.

Notes

Generally, most of the samples worked. I will need to re-do the following wells:

A6, B1, C1, E1 (because there was final low volume), and E3.