Table 3 Primer sequences and conditions used to amplify ELP/CTI genes and transcripts

FT dunnart transcript

FT_ELP_F

GTCAAGTGTTATCTACTGGCAGCACCATG

488

94°C for 2 min; 35 cycles of 94°C for 30 sec; 59°C for 30 sec; 68°C for 1 min; 68°C for 10 min

FT_ELP_R

CCCAAAGTGCTGTTAATGCTTTATTGTAGC

Opossum transcript

mELP_NheI_F

GCTAGCAAGGTTTTCTCTCAGTGCCATC

488

94°C for 2 min; 35 cycles of 94°C for 30 sec; 60°C for 30 sec; 68°C for 30 sec; 68°C for 10 min

mELP_BamHI_R

GGATCCTGTTAATGCTTTATTGTACCAG

Tammar transcript

tELP_NheI_F

GCTAGCAAGTGTAGTCTACCAGTGGCACC

479

94°C for 2 min; 35 cycles of 94°C for 30 sec; 58°C for 30 sec; 68°C for 30 sec; 68°C for 10 min

tELP_BamHI_R

GGATCCTGTTAATGCTTTATTGTACCAG

Dog transcript

Dog_ELP_Ex1_F

GCCTAGAACATTCAGCTATTGGCACC

449

94°C for 2 min; 35 cycles of 94°C for 30 sec; 55°C for 30 sec; 68°C for 1 min; 68°C for 10 min

Dog_ELP_Ex3_R

TGAATGTTTTATTGACCTAGACCTGGAGG

Cow transcript

bELP_NheI_F

GCTAGCAACTCACAGCTCCTCACACCATG

463

94°C for 2 min; 35 cycles of 94°C for 30 sec; 58°C for 30 sec; 68°C for 30 sec; 68°C for 10 min

bELP_BamHI_R

GGATCCGAACACTTTATTGACCCAGTCCTG

FT dunnart gene

FT_ELP_F

GTCAAGTGTTATCTACTGGCAGCACCATG

4771

94°C for 2 min; 35 cycles of 94°C for 30 sec; 55°C for 30 sec; 68°C for 6 min; 68°C for 10 min

FT_ELP_R

CCCAAAGTGCTGTTAATGCTTTATTGTAGC

Koala gene

tELP_Ex1_F

GGTAGCAAGTGTAGTCTACCAGTGGCACC

1428

94°C for 2 min; 35 cycles of 94°C for 30 sec; 52°C for 30 sec; 68°C for 4 min; 68°C for 10 min

tELP_BamHI_R

GGATCCTGTTAATGCTTTATTGTACCAG

Tammar gene (6.2 kbpromoter)

T7

TAATACGACTCACTATAGGG

6326

94°C for 2 min; 35 cycles of 94°C for 30 sec; 57°C for 30 sec; 68°C for 8 min; 68°C for 10 min

tELP_Prom_R

GACTGATCAGACCAATATAAGCTT

Tammar gene (7.9 kbpromoter)

T7

TAATACGACTCACTATAGGG

8044

94°C for 2 min; 35 cycles of 94°C for 30 sec; 57°C for 30 sec; 68°C for 8 min; 68°C for 10 min

tELP_Ex1_R

GAGGGCCAACGATGGTAAATTTCAT