PCR 1 amplifying ITS2 marker for Box 1 - O.Fav deep

Protocol

Samples:

• OFAV = Orbicella faveolata
• OFRA = Orbicella franksi
• S = Shallow
• P = Deep

Species	Treatment	ID
OFAV	P	313
OFAV	P	352
OFAV	P	356
OFAV	P	359
OFAV	P	360
OFAV	P	369
OFAV	P	OHS3
OFAV	P	2HS12
OFAV	P	9AZ5
OFAV	P	9AZ9
OFAV	P	9EV35

Master Mix:

Component	Volume for 1 rxn	Volume for 5 rxn (+10% error) in triplicate
2x MyTaq Mix	12.5 uL	495.00 uL
Molecular Water	9.5 uL	376.20 uL
ITS2 Primer Forward (10 uM)	1.0 uL	39.6 uL
ITS2 Primer Reverse (10 uM)	1.0 uL	39.6 uL

Strip Tube Set Up:

Strip (replicate) Number	A	B	C	D	E	F	G	H
1	313	352	356	359	360	369	OHS3	2HS12
2	313	352	356	359	360	369	OHS3	2HS12
3	313	352	356	359	360	369	OHS3	2HS12
4	9AZ5	9AZ9	9EEV35
5	9AZ5	9AZ9	9EEV35
6	9AZ5	9AZ9	9EEV35

Gel Map:

Combined Gel with PCR8

Failed: 352, 356, OHS3, 9AZ5, 9AZ9, 9EV35, 2HS12_2 or H2, 9GT4, 9GT46, 9IR44, 9JQ4, 9JQ53, CX, DW