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Table 2 Primers used in this study

From: The RASSF gene family members RASSF5, RASSF6 and RASSF7 show frequent DNA methylation in neuroblastoma

Gene

Method

Primer sequence

Product (bp)

AT °C

Primer design

RASSF2A

MSP

F: 5′-GTTCGTCGTCGTTTTTTAGGCG-3′

109

62

[13]

  

R: 5′-AAAAACCAACGACCCCCGCG-3′

   

RASSF2A

MSP

F: 5′-AGTTTGTTGTTGTTTTTTAGGTGG-3′

109

60

[13]

  

R: 5′-AAAAAACCAACAACCCCCACA-3′

   

RASSF4

COBRA

F: 5′-AGGATAYGATATATGTAGTGGTTTTTGGATT-3′

270

TD 65-55

[14]

  

R: 5′-ATTATAACCCCTAAATTACTTAACAAAAATACCAAA-3′

   

RASSF5A*

BSP

F: 5′-TTAGGAAAGAGGAATATTTTAT-3′

434

TD 60-50

[12]

  

R: 5′-TAAACCTTCAACCCTACCTCTTTC-3′

   

RASSF5C**

BSP

F: 5′-GGGGTTTAGAGTTAGGGGTTTA-3′

345

TD 60-50

[12]

  

R: 5′-TATAACTTTATCCCTTTACTA-3′

   

RASSF6

COBRA

F: 5′-GTATAGGGAGTGGTTTAGGTTTTTTGATAT-3′

353

TD 67-57

[10]

  

R: 5′-ATCCCCATTTTTTACCTATTATTCACACTATA-3′

   

RASSF7

BSP

F: 5′-GAGAAAAGTTAGGTTTTAGA-3′

592

TD 62-52

[12]

  

R: 5′-CTCAACAACCTTCTAATATAA-3′

   

RASSF8

BSP

F: 5′-TTTTATAATGTAGYGTTGGYGTTTTAGTTT-3′

374

TD 67-57

[10]

  

R: 5′-CRAAACTCRACRAAACTAAACRAAAAACT-3′

   

RASSF10

BSP

F: 5′-TTGTTTTTGTTGTTTTYGTYGTTTTAGTAGATT-3′

634

TD 67-57

[10]

  

R: 5′-CRATTAAACTTAACCAATTTACRAAAAACCTTA-3′

   

RASSF2A

qRT-PCR

F: 5′-AAGGGGTGGAGAGTGATATGAAGAG-3′

194

60

[15]

  

R: 5′-AGGGACGTTTGGTGGCTGTAGT-3′

   

RASSF4

qRT-PCR

F: 5′-GGACTGCGCGATGACTGGAC-3′

126

56

[15]

  

R: 5′-CCGACTTCTGAATGGACTTGCTGT-3′

   

RASSF5

End-point RT-PCR

F: 5′-CCTGGGCATGAAACTGAGTGAAGA-3′

188

56

Manual

  

R: 5′-tgatggcatctaggggcaggtaga-3′

   

RASSF6

End-point RT-PCR

F: 5′-ATGGAGAGACTGAAGATGGC-3′

203

56

[15]

  

R: 5′-CAGGGTGTTGCTGTGATAAG-3′

   

RASSF7

End-point RT-PCR

F: 5′-CAGCAGAGCGAGCCTTGCAGGCTCA-3′

149

59

Manual

  

R: 5′-CTGAGTGCCAGGAGGGCCCCTGTCA-3′

   

RASSF10

qRT-PCR

F: 5′-CCATGACCCAGGAGAAACAG-3′

226

60

[10]

  

R: 5′-TGCTGGCGAATTGTGTGGTC-3′

   
  1. *cg17558126, **cg02589695.
  2. MSP, Methylation specific PCR; BSP, Bisulfite sequencing PCR; COBRA, Combined bisulfite restriction PCR.
  3. qRT-PCR, Quantitative real time PCR; AT, Annealing temperature; TD, Touch down.