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Table 1 Primers and amplification conditions.

From: Methylation alterations are not a major cause of PTTG1 missregulation

Primer

Sequence (5'-3')

Amplicon

Amplification conditions

PTTG MF1

TTTCGGATTGTTAATTGGATTAAC

168 bp

1. [95°C for 7'] 2. [95°C for 0'', 60°C for 20'', 72°C for 25 '']x45 3. Melting curve

PTTG MR1

AAAAACAAAAACTAAACAACGAA

 

2. [95°C for 0'', 60°C for 20'', 72°C for 25 ''] × 45

PTTG UF1

TTGGATTGTTAATTGGATTAATGG

166 bp

3. Melting curve

PTTG UR1

AAAAACAAAAACTAAACAACAAA

  

PTTG MF2

GTTTTATTTGGTGATTACGTTTACG

123 bp

1. [95°C for 7']

PTTG MR2

ACCGCATTCATCTAAAACCG

 

2. [95°C for 0'', 60°C for 20'', 72°C for 25 ''] × 45

PTTG UF2

TTTTATTTGGTGATTATGTTTATGG

125 bp

3. Melting curve

PTTG UR2

ACAACCACATTCATCTAAAACCAC

  

PTTG-1F

AAAGTAGCTACCATTCCTGC

124 bp

1. [95°C for 7'] 2. [95°C for 30'', 60°C for 30'', 72°C for 30 '']x45 3. [72°C for 3']

PTTG-1R

TGCCCTGTAAAAGCAAAAT

 

2. [95°C for 30'', 60°C for 30'', 72°C for 30 ''] × 45

PTTG-1 Anc

FL-AATAGCCCAACATAATAGAATCTATTTT-Ph

 

3. [72°C for 3']

PTTG-1 Sen

GCTACCATTCCTGCCTTAATA-Cy5

  

PTTG-3F

TGCTGACAGGTGCTGGTACT

128 bp

1. [95°C for 7'] 2. [95°C for 30'', 60°C for 30'', 72°C for 30 '']x45 3. [72°C for 3']

PTTG-3R

AAGAAGCCATAATCCTTAGTTTTCA

 

2. [95°C for 30'', 60°C for 30'', 72°C for 30 ''] × 45

PTTG-3 Anc

FL-TTTGAGTCATGCTACTCGAATTACAT-Ph

 

3. [72°C for 3']

PTTG-3 Sen

GGTACTTAAATTTCCGATTTTAAC-Cy5

  

p16 Forward outer

GTAGGTGGGGAGGAGTTTAGTT

283 bp

1. [94°C for 3']

p16 Reverse outer

TCTAATAACCAACCAACCCCTCC

 

2. [94°C for 30'', 50°C for 30'', 72°C for 30''] × 20

3.[72°C for 4']

p16 Forward inner

GGGGGAGATTTAATTTGG

190 bp

1. [94°C for 3'] 2. [94°C for 30'', 50°C for 30'', 72°C for 30''] × 30 3. [72°C for 4']

p16 Reverse inner

CCCTCCTCTTTCTTCCTC

 

2. [94°C for 30'', 50°C for 30'', 72°C for 30''] × 30

3. [72°C for 4']

qPCR PTTG Fw

AGGCACCCGTGTGGTTGCT

126 bp

1. [50°C for 2']

qPCR PTTG Rev

TAAGGCTGGTGGGGCATC

 

2. [95°C for 2']

qPCR RPL13A Fw

CCTGGAGGAGAAGAGGAAAGAGA

125 bp

3. [95°C for 5'', 60°C for 30''] × 40

qPCR RPL13A Rev

TTGAGGACCTCTGTGTATTTGTCAA

 

4. Melting curve