Table 1

Primer list and amplification/sequencing strategies

PRIMER LIST


Gene

Primer

Forward primer (5'-3')

Reverse primer (5'-3')

(bp)


AgAcp34A-1

69 ex

ATTGAACGAGCACCACCGC

ACAATGCAGAACCTTCGAC

735

AgAcp34A-1

69 n

ACGCCAGGCTTGTACTCTC

CACTTATAAACTAGCTACC

632

AgAcp34A-2/3

70 ex

ATTCCCGTAACTATCTTGC

TATAACTCACGGGCGATTC

793

AgAcp34A-2/3

70 n

AATGTGTCCTTTCTGAACC

CTGCCCAATTAACCAATAG

685

AgAcp34A-1/2/3

exo1 (+)

TCGCCCTAGTGGCTGTTG

-

-

AgAcp34A-2/3

3UTR (-)

-

TCGTCCATTCCCATCGCAG

-

AgAcp34A-1/2

exo2a (+)

TAGATAACAGACAGTTACC

-

-

AgAcp34A-1/2

exo2b (+)

ATGCCCATAAATACTTTAG

-

-

AgAcp34A-1/2

exo2a (-)

-

GTCAACAAGCCCTACAAGA

-

AgAcp34A-3

exo2c (+)

ATACCCAAMCTGCCTATGC

-

-

AgAcp34A-3

exo2d (+)

CTATGCGCCGGCAGGTTTC

-

-

AgAcp34A-3

exo2b (-)

-

ATTTTAGAAACCTGCCGGC

-

AgAcp34A-1

69 C (-)

-

TCTATAGAYAGTATCTACG

-

AgAcp34A-2/3

70 C (-)

-

AAATTGTTCATTGAGAGTC

-


PCR STRATEGIES


Nested PCR strategy to selectively amplify AgAcp34A-2 and AgAcp34A-3a


Gene

Portion

Forward primer (5'-3')

Reverse primer (5'-3')

(bp)


AgAcp34A-2

N terminal

70 n (+)*

exo2a (-)

326

C terminal

exo2a (+)

3UTR (-)*

276

AgAcp34A-3

N terminal

70 n (+)*

exo2b (-)

296

C terminal

exo2c (+)

3UTR (-)*

289


Nested RT-PCR strategy to selectively amplify transcripts (cDNA)


Gene

Round

Forward primer (5'-3')

Reverse primer (5'-3')

(bp)


AgAcp34A-1

1° round

exo1 (+)

69 C (-)

260

2° round

exo2a (+)

69 C (-)

170

AgAcp34A-2

1° round

exo1 (+)

70 C (-)

266

2° round

exo2a (+)

70 c (-)

176

AgAcp34A-3

1° round

exo1 (+)

70 C (-)

263

2° round

exo2c (+)

70 C (-)

189


a 1° round using common external primers: 70 ex (+)/(-); * sequencing primers

Mancini et al. BMC Evolutionary Biology 2011 11:292   doi:10.1186/1471-2148-11-292

Open Data