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

Protocol

Notes:

• Samples that were amplified with the previous protocol were re run with new protocol (see ITS2_PCR1_12)
• DNA for sample 9AZ5 was re aliquoted from Diluted PCR stock as it failed all three runs in ITS2_PCR1_12

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	358
OFAV	P	359
OFAV	P	360
OFAV	P	369
OFAV	P	OHS3
OFAV	P	362
OFAV	P	365
OFAV	P	366
OFAV	P	367
OFAV	P	368
OFAV	P	310
OFAV	P	357
OFAV	P	371
OFAV	P	Blank
OFAV	P	9AZ5
OFAV	P	OHS4
OFAV	P	2HS2
OFAV	P	2HS6
OFAV	P	9GT46
OFAV	P	CX
OFAV	P	DW

Master Mix:

Component	Volume for 1 rxn	Volume for 24 rxns (+10% error)
2x MyTaq Mix	25.0 uL	660 uL
Molecular Water	18.0 uL	475.2 uL
ITS2 Primer Forward (10 uM)	2.0 uL	52.8 uL
ITS2 Primer Reverse (10 uM)	2.0 uL	52.8 uL
DNA	3.0 uL	NA

Strip Tube Set Up:

Strip (replicate) Number	A	B	C	D	E	F	G	H
1	313	352	356	358	359	360	369	OOHS3
2	313	352	356	358	359	360	369	OOHS3
3	313	352	356	358	359	360	369	OOHS3
4	362	365	366	367	368	310	357	371
5	362	365	366	367	368	310	357	371
6	362	365	366	367	368	310	357	371
7	blank	9AZ5	OHS4	2HS2	2HS6		CX	DW
8	blank	9AZ5	OHS4	2HS2	2HS6		CX	DW
9	blank	9AZ5	OHS4	2HS2	2HS6		CX	DW

Gel Map:

Failed: 352, 356, 360, 369, OHS3, 9AZ5, 9GT46, DW