4. Antibiotics: kanamycin and rifampicin.
5. Liquid nitrogen.
6. Incubators (37 and 28
C).
2.3 Arabidopsis
Transformation
1. T-DNA transformed Agrobacterium.
2. LB liquid medium.
Table 1
Sequence of primers used in this study
Primer name
Primer sequence (5
0 - 3
0 )
Forward oligo of sgRNA
GATTGNNNNNNNNNNNNNNNNNNNN
Reverse oligo of sgRNA
AAACNNNNNNNNNNNNNNNNNNNNC
M13-F
TGTAAAACGACGGCCAGT
M13-R
CAGGAAACAGCTATGACC
pCAMBIA1300-P1
CCAGGCTTTACACTTTATGC
pCAMBIA1300-P2
GCGATTAAGTTGGGTAACGC
AtU6-26-sgRNA forward
CGTTGAACAACGGAAACTCGAC
AtU6-26-sgRNA reverse
AAAAAAAGCACCGACTCGGT
GL2 genotyping forward
AGTTAGGGTTCAGTTGCATG
GL2 genotyping reverse
AGTTATAGTAGCTGGTAACAG
ROS1-GFP-5 arm forward
GAATTCGTTATTCAACAGGTGAGACGGCTGATTC
ROS1-GFP-5 arm reverse
TGCTCACCATGGCGAGGTTAGCTTGTTGTCC
ROS1-GFP-GFP forward
TAACCTCGCCATGGTGAGCAAGGGCGAGGAG
ROS1-GFP-GFP reverse
TTTGCTTGCCTTACTTGTACAGCTCGTCCATGCCGT
ROS1-GFP-3 arm forward
GTACAAGTAAGGCAAGCAAACAAATACAAGCTTATG
ROS1-GFP-3 arm reverse
GACAACAATAGAATTCTTTGGTCCGGTTGAACTATC
DME-D1633A-5 arm forward
GGAACTTGTATTGATGATATGAAGGTTGACACG
DME-D1633A-5 arm reverse
GACATGCATTACAgTTagcTCTACTCTTTGTGC
DME-D1633A-3 arm forward
GCACAAAGAGTAGAgctAAcTGTAATGCATGTC
DME-D1633A-3 arm reverse
CCGTAGAGTCATTCCAAACTGTTCAATGTTG
pCAMBIA3301-foward
CTTCCGGCTCGTATGTTGTG
pCAMBIA3301-reverse
CTCTTCGCTATTACGCCAGC
ROS1-GFP-specific forward
GCAGTTGGAAAAGAGAGAACCTGATGATCC
ROS1-GFP-specific reverse
CTGAACTTGTGGCCGTTCACGTC
ROS1-GFP external forward
ACCTGATGATCCATGTTCTTATTTG
ROS1-GFP external reverse
CCTTGTACAACTCTAGGACTGTT
CRISPR/Cas9-Based Genome Editing in Arabidopsis
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