Table 1

nucleotide sequences of primers and the PCR products size

TARGET

sequences (primer 3 design) or Qiagen Geneglobe reference number

NCBI reference gene number

NCBI detected transcript number

lenght of amplicon

amplified exons*


    GAPDH (fragments)

4141

NM_002046

    GAPDH 5'end

GAPDH 5' Forward

5'-GAAGGTGAAGGTCGGAGT-3'

252 bp

2/3

GAPDH 5' Reverse

5'-GAAGATGGTGATGGGATTTC-3'

    GAPDH 3' end

GAPDH 3' Forward

5'-AAACCTGCCAAATATGATGACAT-3'

228 bp

8/9

GAPDH 3' Reverse

5'-ACCCTGTTGCTGTAGCCAAA-3'

FASN (fragments)

2194

NM_004104

    FASN 5' end

FASN 5' Forward

5'-ATCCGCTCGTTGTACCAGTC-3'

281 bp

7

FASN 5' Reverse

5'-GATCTCAGGGTTGGGGCTAT-3'

9

    FASN 3' end

FASN 3' Forward

5'-GGTCTTGAGAGATGGCTTGC-3'

197 bp

34/35

FASN 3' Reverse

5'-TTGGCAAAGCCGTAGTTG-3'

36

    House keeping genes

ALAS1 (Aminolevulinate delta synthase 1)

QT00073122

211

NM_000688

113 pb

9/10

B2M (beta2 microglobulin)

QT00088935

567

NM_004048

98 pb

1/2

Cyclophilin A (peptidylprolyl isomerase)

Forward

5'-TGCTGACTGTGGACAACTCGA-3'

5478

NM_203431

212 bp

4

Reverse

5'-TCATAATCATAAACTTAACTCTGCA ATCC-3'

    other genes

TNFalpha(Tumor necrosis factor)

QT01079561

7124

NM_000594

104 bp

1/2/3

COX 2 (cyclooxygenase 2)

QT00040586

5743

NM_000963

68 bp

1/2


Detail of primers sets specified for RT-PCR analysis

Some primers are designed by Qiagen manufacturer (Gene globe Quantitect primer assay products). Others are designed using Primer 3 software.

(*) primer sets are designed to cross exon/intron boundaries in order to prevent coamplification of genomic DNA.

Fajardy et al. BMC Molecular Biology 2009 10:21   doi:10.1186/1471-2199-10-21

Open Data