Project: Thermal Transplant Molecular

Goal

To test ITS2 sequencing prep on 4 Porites astreoides samples and 1 Pocillopora spp. sample from D. Becker. 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

Samples

Position	Vial #	Life Stage	Species	DNA conc. (ng/uL)	DNA for Dilution (uL)	Water Vol (uL)	Total Vol (uL)	Final DNA Conc. (ng/uL)
1	L-1081	Larvae	P. astreoides	46.2	1.1	18.9	20	2.5
2	L-1084	Larvae	P. astreoides	46.4	1.1	18.9	20	2.5
3	18-466	Adult	P. astreoides	21.2	2.4	17.6	20	2.5
4	18-44	Adult	P. astreoides	27.2	1.8	18.2	20	2.5
5	E1	Adult	Pocillopora spp.	6.06	8.3	11.7	20	2.5
6	NEG	NA	NA	0	0	20	20	0

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.

Master Mix Calculation

Strip Tube planning

Gel Results

Gel Electrophoresis

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