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Table 1 Primers used for RT-PCR analysis of CUL family members

From: Analysis of CUL-5 expression in breast epithelial cells, breast cancer cell lines, normal tissues and tumor tissues

Primer Sequences

Primer Location

and GenBank Accession #

Annealing

Temp. and

Predicted

product size

Restriction

endonuclease

and cut site

CUL-1 forward:

5'-CGCTGGCTTTGTGGCTGCTC-3'

CUL-1 reverse:

5'-TGTGGCGGCTGGCGTAGAA-3'

nt 1510 – 1529

nt 2051 – 2069

XM027270

58°C

560-bp

Bgl II

1891

CUL-2 forward:

5'-CAGCGAAAGGGATGACAGAGAATG-3'

CUL-2 reverse:

5'-TGAGTAAGAGGCCACGCACCA-3'

nt 1351 – 1374

nt 1688 – 1708

XM005814.3

54°C

358-bp

Aat II

1435

CUL-3 forward:

5'-AGAGCGGAAAGGAGAAGTCGTAGA-3'

CUL-3 reverse:

5'-CTCAAAGTCACCCGCAATAGTT-3'

nt 833 – 856

nt 1424 – 1445

XM051985.1

53°C

613-bp

EcoR I

1160

CUL-4A forward:

5'-AACTCCACGCTGCCCTCCATCTG-3'

CUL-4A reverse:

5'-TGCTGCCCGCCCCTCACC-3'

nt 346 – 368

nt 840 – 857

XM027482.1

58°C

512-bp

Sau3A I

452

CUL-4B forward:

5'-GCGGAGCTGCTTTCACCA-3'

CUL-4B reverse:

5'-TTCTTCAACCGTTTCTTTCATCTG-3'

nt 1400 – 1417

nt 1969 – 1992

NM003588.1

54°C

593-bp

Hae III

1551

CUL-5 forward:

5'-GAGTGGCTAAGAGAAGTTGGTATG-3'

CUL-5 reverse:

5'-TCTTCTCTCATCCTTTCTGTAGTG-3'

nt 1521 – 1544

nt 2171 – 2194

NM003478.1

58°C

674-bp

EcoR I

1767

Ribosomal S9 forward:

5'-GCAGGCGCAGACGGGGAAGC-3'

Ribosomal S9 reverse:

5'-CGCGAGCGTGGTGGATGGAC-3'

nt 6 – 25

nt 397 – 416

NM001013.2

62°C

411-bp

Dde I

349

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