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Table 2 Primers used in this study

From: Pseudo attP sites in favor of transgene integration and expression in cultured porcine cells identified by streptomyces phage phiC31 integrase

Name

Sequence

Comments

attB-F

GTCATTAATCGCCATTCAGGCTGCGCA

To amplify 400 bp attB site in AseI of pEGFP-N1

attB-R

GTCATTAATCTCGGCCTCGACTCTAG

attL-F

GGCGAGAAAGGAAGGGAAG

To detect the attL site, 400 bp

attL-R

ATTAACCCTCACTAAAGGGA

attR-F

TCAAAGTAAACGACATGGTG

To detect the attR site,520 bp

attR-R

CCAACCGCTGTTTGGTCTGC

IV-R1

GCTTATAGATACCGTAGACAT

TAIL-PCR to identify integration site

IV-R2

TCCCGTGCTCACCGTGACC

IV-R3

ATCAACTACCGCCACCTCG

IV-F2

CGCCACCTCTGACTTGAGCG

 

5113-LF

GTTGGGATTTGGCTCCAGCC

Use with IV-R1 for determination of 5′ end of 5113 pseudo attP site

5113-RR

TCTTAACACCCCATTGTTCTC

Use with IV-F2 for determination of 3′ end of 5113 pseudo attP site

5156-LF

GCCTTAATGAGGGCCAGAGC

Use with IV-R1 for determination of 5′ end of 5156 pseudo attP site

5156-RR

GTTGACCCATTTTCCTGCTCTTCG

Use with IV-F2 for determination of 3′ end of 5156 pseudo attP site

1015-LF

GTCCTGCCCTGACCCTTTGG

Use with IV-R1 for determination of 5′ end of 1015 pseudo attP site

1015-RR

GCCAACCTCCAAGTTGTTGG

Use with IV-F2 for determination of 3′ end of 1015 pseudo attP site

2015-LF

GCAAGGGGTT TTGAAACTGC

Use with IV-R1 for determination of 5′ end of 2015 pseudo attP site

2015-RR

AAAAGTGCCTCAGAACACCAC

Use with IV-F2 for determination of 3′ end of 2015 pseudo attP site

5113-pF

TCAAAGTAAACGACATGGTGGAGTGTAGCAGTTCCTAGGC

ABI-REC cloning of 5113 pseudo attP site in pBCPB+ plasmid

5113-pR

ATTAACCCTCACTAAAGGGATCTTAACACCCCATTGTTCTC

5156-pF

TCAAAGTAAACGACATGGTGGACCTCTGCTGTGCCCAGAG

ABI-REC cloning of 5156 pseudo attP site in pBCPB+ plasmid

5156-pR

ATTAACCCTCACTAAAGGGACCAACCACTGGCTTCCCTCG

1015-pF

TCAAAGTAAACGACATGGTGACCCCCACCCTGAGCTACTC

ABI-REC cloning of 1015 pseudo attP site in pBCPB+ plasmid

1015-pR

ATTAACCCTCACTAAAGGGAACTCCCCAGCAACCAGCTTC

2015-pF

TCAAAGTAAACGACATGGTGATGAGACAAGAGGGACCAGG

ABI-REC cloning of 2015 pseudo attP site in pBCPB+ plasmid

2015-pR

ATTAACCCTCACTAAAGGGAAAAAGTGCCTCAGAACACCAC

Pseudo-P1R

CACCATGTCGTTTACTTTGA

ABI-REC cloning of pig pseudo site

BCPB-S

GCTGCAAGGCGATTAAGTTGGG

Sequencing primer

EGFP-QF

TGAACCGCATCGAGCTGAAGGG

Real-time PCR primers, 130 bp

EGFP-QR

ACCTTGATGCCGTTCTTCTGCTTG

TFRC-QF

GAGACAGAAACTTTCGAAGC

Real-time PCR primers, 81 bp

TFRC-QR

GAAGTCTGTGGTATCCAATCC