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International Journal of Marine Science 2014, Vol.4, No.50, 1-22
http://ijms.biopublisher.ca
12
Table 4
.
PCR primers, mastermix composition, and thermocycling conditions
. When two values are given under the primer concentration (“nM primer”), annealing temperature,
or PCR cycles (“Cycle #”) columns, they correspond to values used in the
Seriatopora hystrix
elevated (i.e., ETE) and variable (i.e., VTE) temperature experiments, respectively. When only one
value appears, the same conditions were used for samples of both experiments. Full genes names have been written out in the text and Tables A1-3.
Note:
a
“bp” = base pairs.
b
ETE only; VTE assays were conducted by Mayfield et al. (2012a).
c
VTE only; ETE assays were conducted by Mayfield et al. (2011).
Gene
Length
(bp)
a
Sequence (‘5-3’)
nM
primer
Annealing
temp. (
℃
)
Cycle # Cellular process
NCBI
accession
Reference
Solaris spike
NA
TGCAAAGCCAATTCCCGAAG
CCATTGTAGTGAACAGTAGGAC
125
63
36
34
exogenous spike NA
Putnam et al., 2013
SYMBIODINIUM
TARGET GENES
rbcL
126
CAGTGAACGTGGAGGACATGT
AGTAGCACGCCTCACCGAAA
200
b
60.5
32
photosynthesis
AAG37859
Mayfield et al., 2012a
psI
136
GTGGAGTTGACATTGACTTGGA
TGCTGCTTGGTGGTCTTGTA
500
b
59
39
photosynthesis
HM156699
Mayfield et al., 2012a
pgpase
100
TGACAAACAATTCCACCAAGAG
GCTGCAAAGGATGATGAGAAG
250
b
60
32
photosynthesis
EU924267
Crawley et al., 2010
nrt2
97
CCACCCATTTCAGGACCTAT
CCAGGGACCTAGCAAACAA
200
61.5
62
33
38
metabolism
HM147134
Mayfield et al., 2013a
hsp70
86
CTGTCCATGGGCCTGGAGACT
GTGAACGTCTGTGCCTTCTTGGTT
500
c
62.5
40
stress response
EU476880
Mayfield et al., 2009
apx1
107
GCCAAGTTCAAGGAGCATGTA
AGCTGACCACATCCCAACT
150
b
61
40
stress response
HM156698
Mayfield et al., 2012a
SERIATOPORA HYSTRIX
TARGET GENES
actb
100
GACTCGTATGTCGGTGATGAAGCT
TTTCCATGTCATCCCAGTTGGT
50
60
29
cytoskeleton
HM147127
Mayfield et al., 2010
trp1
108
GCTGCAAAGATCGCTAACCTG
TCCAACCAAGGTGACCTCTTC
50
61.5
33
cytoskeleton
HM147128
Mayfield et al., 2010
tuba
110
ACATCTGTCGGCGCAACCT
GCACCATCAAAGCGCAGTGA
50
61.5
33
cytoskeleton
HM147129
Mayfield et al., 2010
ezrin
162
CAGCGGAAAATGCTTCAGC
TTCATCAACCCTCTGCTTAGTG
400
59
35
cytoskeleton
HM147133
present study
cplap2
174
GCAAACGGACGGAATCAAAGA
GGCATCTTCAAACACGGAGA
100
60
35
osmoregulation
HM156700
present study
oatp
130
ACGGTTGGCTGTTTGACACT
AACACATGGCAAGTTCCGG
250
200
60
35
osmoregulation
HM147131
present study
trcc
114
TGGTCGGTTTGGCTGTTG
TCCATTGTGGGAGAGATTCTTC
250
60
30
osmoregulation
HM147132
present study
hsp70
62
AATCCAGGAGTTGCTCAGAAATT
TCTGACACAGCTTCACTCTTA
500
c
59
28
stress response
AB201749
Mayfield et al., 2011