ITS2_Box1_PCR1_9

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 352
OFAV P 356
OFAV P OHS3
OFAV P 9AZ5
OFAV P 9EV35
OFAV P 9GT4
OFAV P 9GT46
OFAV P 9IR44
OFAV P 9JQ4
OFAV P 9JQ53
OFAV P CX
OFAV P DW

Master Mix:

Component Volume for 1 rxn Volume for 5 rxn (+1% 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
1 352 356 OHS3 9AZ5 9EV35 9GT4 9GT46 9IR44
2 352 356 OHS3 9AZ5 9EV35 9GT4 9GT46 9IR44
3 352 356 OHS3 9AZ5 9EV35 9GT4 9GT46 9IR44
4 9JQ4 9JQ53 CX DW        
5 9JQ4 9JQ53 CX DW        
5 9JQ4 9JQ53 CX DW        

Gel Map:

Failed: All samples – next steps are re diluting primers and MyTaq to rid potential contamination

Written on June 6, 2022