Project: Thermal Transplant Molecular

Goal

Re-doing ITS2 sequencing prep on Porites astreoides adult and larval samples that had low quantities from the 20210302 prep. 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

Samples and Strip Tube Layout

I took the diluted samples from the dilution plate I prepped on 20210302.

Dilution Plate Well	Sample ID	Strip tube position
-	Negative Control (Ultra pure H2O)	1
A6	L-728	2
B1	L-859	3
C1	L-1266	4
E1	18-250	5
E3	L-704	6
E9	E1 (Positive control; Becker)	7

Gel Results

Gel Electrophoresis

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