From: Rapid, high efficiency virus-mediated mutant complementation and gene silencing in Antirrhinum
Purpose | Primer name | Sequence (5′-3′) | Notes |
---|---|---|---|
VIGS | AscI-PDS-F | TGGCGCGCCCAATGAGCCTTACCGTGCAT | Adds Asc I sites to AmPDS fragment |
AscI-PDS-R | TGGCGCGCCGTAGTGCTTCAGAGGATCTTACAA | ||
PDS-R | GTAGTGCTTCAGAGGATCTTAC | First-round AmPDS amplification | |
AscI-DIV-R | AGGCGCGCCTACCCCATTCTAAGGTAAAAGG | Adds Asc I site to Div fragment | |
PDS-DIVfusion-F | gtaagatcctctgaagcactacCATATTTTTCCAGCTCAAGCTGG | Adds PDS-R to Div fragment | |
AscI-Hutr-F | AGGCGCGCCATAATACAAGTTGAGCAACAGCG | Adds Asc I site to H 3′-UTR fragment | |
PDS-Hutrfusion-R | gtaagatcctctgaagcactacACAGAGTGATACGCCTCGAT | Adds PDS-R to H 3′-UTR fragment | |
H expression | AscI-Hairy-F | AGGCGCGCCATGCAGTACGACGCAGAACC | Adds Asc I site 5′ to the H ORF |
PDS-Hairyfusion-R | gtaagatcctctgaagcactacTTAGAGCCAAAGAGCACCAGC | Adds PDS-R 3′ to the Hairy ORF | |
AscI-SynH-F | AGGCGCGCCAACAATGCAATATGATGCTGAGCCT | Adds Asc I upstream of SynH ORF | |
FLAG-SynHfusion-R | CTACTTGTCGTCATCGTCTTTGTAGTCCAACCACAATGCTCCTGCT | Replaces H stop with FLAG-tag | |
FLAG-PDSfusion-R | GACTACAAAGACGATGACGACAAGTAGgtagtgcttcagaggatcttac | Adds FLAG to AmPDS fragment | |
RT-PCR | AmPDS-qPCR-F | TCTTTGTAATGGACGGCAAG | RT-PCR for AmPDS |
AmPDS-qPCR-R | ACTTGCCAAACTCTTCCCTG | ||
TRV-CP-F | TGGGTTACTAGCGGCACTGA | RT-PCR for TRV2 RNA | |
TRV-CP-R | GCTCGTCTCTTGAACGCTGA | ||
AmUbi-qPCR-F | CCGAACCATCAGACAAACAAAC | RT-PCR for AmUBI control | |
AmUbi-qPCR-R | TACCCTGGCCGACTACAATA | ||
AmACT-F | CTTGGCCGTCTCCATTTCTT | RT-PCR for AmACT control | |
AmACT-R | TCCTCACAGAGCGTGGATATAG | ||
H-qPCR-F2 | ACCATCCACAACACTCTAATC | RT-PCR for H | |
H-qPCR-R | TGAATATCACCACCGGATTCTC | ||
Div-F | GGGTAGTGGTCATGGATTCG | RT-PCR for Div | |
Div-R | GGAAAGGAAGTTTGTGAATGGAG |