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Table 1 PCR primers

From: Enhancement of pacing function by HCN4 overexpression in human pluripotent stem cell-derived cardiomyocytes

Name

 

Sequence

Product size (bp)

Annealing temperature (℃)

Subcloning PCR

PGK-Neo

Forward

TGGGGCGGGATCTGTAGGGCGCAGTA

2886

64

Reverse

AACTAGATAACTTCGTATAGCATACATTATACGAAGT

  

AAVS1 homology arm/AmpR

Forward

ACCTGCAGACTAGGGACAGGATTGGT

5624

64

Reverse

ACAGATCCCGCCCCACTGTGGGGTGGA

  

CAG-IRES-EGFP

Forward

CGAAGTTATCTAGTTATTAATAGTAATCAATTACGGGG

3614

62

Reverse

CCCTAGTCTGCAGGTCGAGGGATCT

  

Human HCN4

Forward

CAAAGAATTCCTCGAGACCATGGACAAGCTGCCGCCGTC

3647

64

Reverse

CCGCGATATCCTCGAGTCATAGATTGGATGGCAGTTTGGAGC

  

Flanking PCR

Primer 1

 

TGGGCTTTGCCACCCTATGCTGACA

 

64

Primer 2

 

ACAAGCAGAAGAACGGCATCAAGGTGAA

  

Primer 3

 

ACCAACCATCCCTGTTTTCCTAGGACTGA

  

qPCR

GAPDH

Forward

CAACGACCACTTTGTCAAGCTC

144

60

Reverse

TCTCTTCCTCTTGTGCTCTTGC

  

HCN4

Forward

GGTGTCCATCAACAACATGG

66

60

Reverse

GCCTTGAAGAGCGCGTAG

  

MYL2

Forward

GTGCTGAAGGCTGATTACGTTC

121

60

Reverse

TGTAGTCCAAGTTGCCAGTCAC

  

NKX2-5

Forward

GTCCCCTGGATTTTGCATTCAC

100

60

Reverse

ATAATCGCCGCCACAAACTCTC

  

NR2F2

Forward

TCGCCTTTATGGACCACATACG

149

60

Reverse

TTCCACATGGGCTACATCAGAG

  

SCN5A

Forward

AGAAGATGGTCCCAGAGCAATG

131

60

Reverse

AATCTGCTTCAGAACCCAGGTC

  

SHOX2

Forward

ATCGCAAAGAGGATGCGAAAGGGA

86

60

Reverse

TTCCAGGGTGAAATTGGTCCGACT

  

TBX3

Forward

TTGAAGACCATGGAGCCCGAAGAA

91

60

Reverse

CCCGCTTGTGAAACTGATCCCAAA

  

TBX18

Forward

TTAACCTTGTCCGTCTGCCTGAGT

147

60

Reverse

GTAATGGGCTTTGGCCTTTGCACT

  

TNNT2

Forward

TTCACCAAAGATCTGCTCCTCGCT

166

60

Reverse

TTATTACTGGTGTGGAGTGGGTGTGG

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