Pfx platinum taq

Genomic DNA was extracted from Arabidopsis thaliana Columbia-0 (Col-0) seedling tissue [40 ]. Upstream regulatory regions for use in tissue-specific expression vectors were PCR-amplified from Col-0 genomic DNA using Pfx Platinum Taq (Life Technologies) and the following primer pairs: ADH1-EcoRI (5′-GAATTCCACACTGAAGAAAAAGATTACACC-3′) and ADH1-XhoI (5′-CTCGAGCAACAGTGAAGAACTTGCTTTTG-3′); CAB1-NruI (5′-TCGCGAGACTAACTTGTGAGTGAGAGTG-3′) and CAB1-XhoI (5′-CTCGAGGAGGTTGAGTAGTGCAGCAC-3′); COBL1-NruI (5′-TCGCGACTCATGTTTGGTTGTACTACTG-3′) and COBL1-XhoI (5′-CTCGAGCTGAAGCAAAAAAAGAGAGAG-3′); EXP7-EcoRI (5′-GAATTCCGTCAAGGCTGGATATGCTGTG-3′) and EXP7-XhoI (5′-CTCGAGGCTGCGATCTAACAATTTCAGAC-3′); LBD1-EcoRI (5′-GAATTCGCGGAAGAACTTATAAAATAAC-3′) and LBD1-XhoI (5′-CTCGAGCGGCGAAACGAACAAAAAAGTG-3′); SCR-EcoRI (5′-GAATTCGATTGTGATCCTCTGCAACAAAGC-3′) and SCR-XhoI (5′-CTCGAGGGAGATTGAAGGGTTGTTGGTCG-3′); UBQ10-AatII (5′-GACGTCGTATGATCGCGAAGCACCCACCCTAAGC-3′) and UBQ10-XhoI (5′-CTCGAGGACAAATTCGATCGCACAAAC-3′); and WOL1-AatII (5′-GACGTCCTCACACACCACACCATCATTATC-3′) and WOL1-XhoI (5′-CTCGAGCACTTCAAATGTAGGTATTCC-3′). The resulting PCR products were captured into the pCR4 vector (Life Technologies) to create pCR4-ADH1p, pCR4-CAB1p, pCR4-COBL1p, pCR4-EXP7p, pCR4-LBD16p, pCR4-SCRp, pCR4-UBQ10p, and pCR4-WOL1p. All constructs were sequenced (GeneWiz, Inc.) to confirm error-free clones.