ITS2_Box1_PCR1_6

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 370
OFAV P 2HS48
OFAV P 9AZ2
OFAV P 9AZ5
OFAV P 9AZ9
OFAV P 9EV35
OFAV P 9EV4
OFAV P 9EV40
OFAV P 9EV5
OFAV P 9GT35
OFAV P 9GT46
OFAV P 9IR44
OFAV P 9JQ4
OFAV P 9JQ42
OFAV P 9JQ431
OFAV P 9JQ53
OFAV P AZ
OFAV P CX
OFAV P DW
OFAV P 369
OFAV P 371
OFAV P OHS3
OFAV P OHS4
OFAV P 2HS12
OFAV P 352

Master Mix:

Component Volume for 1 rxn Volume for 5 rxn (+10% error) in triplicate
2x MyTaq Mix 12.5 uL 866.25 uL
Molecular Water 9.5 uL 658.35 uL
ITS2 Primer Forward (10 uM) 1.0 uL 69.3 uL
ITS2 Primer Reverse (10 uM) 1.0 uL 69.3 uL

Strip Tube Set Up:

Strip (replicate) Number A B C D E F G H
1 370 2HS48 9AZ2 9AZ5 9AZ9 9EV35 9EV4 9EV40
2 370 2HS48 9AZ2 9AZ5 9AZ9 9EV35 9EV4 9EV40
3 370 2HS48 9AZ2 9AZ5 9AZ9 9EV35 9EV4 9EV40
4 9EV5 9GT35 9GT46 9IR44 9JQ4 9JQ42 9JQ431 9JQ53
5 9EV5 9GT35 9GT46 9IR44 9JQ4 9JQ42 9JQ431 9JQ53
6 9EV5 9GT35 9GT46 9IR44 9JQ4 9JQ42 9JQ431 9JQ53
7 AZ CX DW 369 371 OHS3 OHS4 2HS12
8 AZ CX DW 369 371 OHS3 OHS4 2HS12
9 AZ CX DW 369 371 OHS3 OHS4 2HS12
10 352 352 352          

Gel Map:

No Gel photo – all samples failed Multiple freezing and thawing due to illness resulted in DNA degradation

Written on April 10, 2022