Cloning primers | ||
AGI Number of target and primer direction | Purpose | Primer sequence (5′- -3′) |
At2g31390 – forward | FRK2 bacterial/plant expression | GGGGACAAGTTTGTACAAAAAAGCAGGCTTGATGGCATCCAACGGTGATAAAGG |
At2g31390 – reverse | FRK2 bacterial expression | GGGGACCACTTTGTACAAGAAAGCTGGGTCCTATTTCTTTTCGAG |
At1g66430 – forward | FRK3 bacterial expression/removal of cTP | GGGGACAAGTTTGTACAAAAAAGCAGGCTTGTCTAATCTCAAAGGAAGAGC |
At1g66430 – reverse | FRK3 bacterial expression | GGGGACCACTTTGTACAAGAAAGCTGGGTCTTAAACGACGGCTTTGAG |
At4g10260 – forward | FRK4 bacterial/plant expression | GGGGACAAGTTTGTACAAAAAAGCAGGCTTGATGGCTAATACTCCATTG |
At4g10260 – reverse | FRK4 bacterial expression | GGGGACCACTTTGTACAAGAAAGCTGGGTCCTATTTAGACTTAGATTTC |
At1g06030 – forward | FRK6 bacterial/plant expression | GGGGACAAGTTTGTACAAAAAAGCAGGCTTGATGACGTCATCCAACGGCG |
At1g06030 – reverse | FRK6 bacterial expression | GGGGACCACTTTGTACAAGAAAGCTGGGTCCTATTCTACTTGTATCTTAAG |
At3g59480 – forward | FRK7 bacterial/plant expression | GGGGACAAGTTTGTACAAAAAAGCAGGCTTGATGGCTGCATCTAACGGCGAG |
At3g59480 – reverse | FRK7 bacterial expression | GGGGACCACTTTGTACAAGAAAGCTGGGTCTTAGTTTCCTTTCAGGAGGC |
At5g51830 – forward | FRK1 plant expression | GGGGACAAGTTTGTACAAAAAAGCAGGCTTGATGGGTGAGGATGCAATCTC |
At5g51830 – reverse | FRK1 plant expression stop codon mutation | GGGGACCACTTTGTACAAGAAAGCTGGGTCTGACGATCGAGTAGAAGAAAG |
At2g31390 – reverse | FRK2 plant expression stop codon mutation | GGGGACCACTTTGTACAAGAAAGCTGGGTCCCATTTCTTTTCGAG |
At1g66430 – forward | FRK3 plant expression | GGGGACAAGTTTGTACAAAAAAGCAGGCTTGATGGCTCTCCAAGCCACTAC |
At1g66430 – reverse | FRK3 plant expression stop codon mutation | GGGGACCACTTTGTACAAGAAAGCTGGGTCTGAAACGACGGCTTTGAG |
At4g10260 – reverse | FRK4 plant expression stop codon mutation | GGGGACCACTTTGTACAAGAAAGCTGGGTCCGATTTAGACTTAGATTTC |
At1g06020 – forward | FRK5 plant expression | GGGGACAAGTTTGTACAAAAAAGCAGGCTTGATGGCATCATCCACCGGCG |
At1g06020 – reverse | FRK5 plant expression stop codon mutation | GGGGACCACTTTGTACAAGAAAGCTGGGTCTGAACAAGGgctAGCCGTACAACACC |
At1g06030 – reverse | FRK6 plant expression stop codon mutation | GGGGACCACTTTGTACAAGAAAGCTGGGTCCGATTCTACTTGTATCTTAAG |
At3g59480 – reverse | FRK7 plant expression stop codon mutation | GGGGACCACTTTGTACAAGAAAGCTGGGTCTGAGTTTCCTTTCAGGAGGC |
Primers for analysis of T-DNA insertional mutants | ||
Arabidopsis T-DNA line | Primer target | Primer sequence (5′- -3′) |
frk1–1 (SALK_046463) | gDNA | CTGGGATGAGATTCGACCAT |
gDNA/T-DNA | GGCAAGGTTCCTCAATCAAA | |
cDNA flanking insert | ATGTTAGCTGATATTCTAAG | |
cDNA flanking insert | TAGCAGCTTCTTCTGATGGC | |
frk2–1 (SALKseq_17726) | gDNA/T-DNA | TCATGGCGAGGATCTTTTGCT |
gDNA | CACACTAGTTTTGCCTTCTGGT | |
cDNA flanking insert | TAGTGGAGCCGTGTAGGTCA | |
cDNA flanking insert | AGCTCAACATCGCTCACCTT | |
frk2–2 (SALK_114786) | gDNA 5’ | TTCTTCTCCTTGTCCTTCCTT |
gDNA/ T-DNA 3’ | CGAGACGAGAAACAGCGATT | |
cDNA flanking insert | CTTCGGCGAGATGCTAATCG | |
cDNA flanking insert | TCCCAGCCAACATATGACCG | |
frk3–1 (SALK_044085) | gDNA | CGTCCTATCTCCATCAAAGC |
gDNA/T-DNA | CGCTTGTCGTCGGTACAA | |
cDNA flanking insert | GTTTTACCGAAACCCGAGTG | |
cDNA flanking insert | CCGCCTTAGCTGCAGAAATA | |
frk3–2 (SALK_035386) | gDNA | CCGCCTTAGCTGCAGAAATA |
gDNA/T-DNA | GTTTTACCGAAACCCGAGTG | |
cDNA flanking insert | GTTTTACCGAAACCCGAGTG | |
cDNA flanking insert | CCGCCTTAGCTGCAGAAATA | |
frk5–1 (SALK_027635) | gDNA | CGCTTTTGTCACTTTGCGTTCTGAC |
frk5–2 (SALK_057002) | gDNA/T-DNA | CGAGTACGGACTGGTCATCAACAA |
cDNA flanking insert | GATGATTTCGGTCATATGCTCGC | |
cDNA flanking insert | GGAGAAGCCGTACAACACCA | |
frk7–1 (GABI_253H07) | gDNA | CAGCACGGTCATCAACAATC |
gDNA/T-DNA | TAGCGAACGGTGCTACATCA | |
T-DNA (GABI) | ATATTGACCATCATACTCATTGC | |
cDNA flanking insert | AACGGTGTCTCTGCTGAAGG | |
cDNA flanking insert | TCAGGAGGCTCTGAACTTCA | |
SALK T-DNA Left border | T-DNA in SALK lines | TGGTTCACGTAGTGGGCCATCG |
Positive control gene (At1g49350) | cDNA | GCGTGGATGCCGTTGAAAAT |