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Supplemental Material for Zhang et al., 2018

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posted on 2018-04-27, 14:59 authored by Xiao-Ran Zhang, Jia-Bei He, Yi-Zheng Wang, Li-Lin Du
Figure S1 shows that a donor-free transformation using a gene-targeting sgRNA insert resulted in a substantially lower transformation efficiency than a control transformation using a control sgRNA insert examples of spindle categories.
Figure S2 shows the effects of different amounts of the gapped plasmid and the sgRNA insert on rpl42-P56Q knock-in when using the split-ura4 system.
Figure S3 shows the effects of different types of donor DNA on rpl42-P56Q knock-in when using the split-ura4 system.
Figure S4 shows the effects of different amounts of donor DNA on rpl42-P56Q knock-in when using the split-ura4 system.
Figure S5 shows that tor2-L2048S mutants generated using the split-ura4 system behaved as expected.
Figure S6 shows that cdc25-C532Y mutants generated using the split-ura4 system behaved as expected.
Figure S7 shows that mECitrine-Ypt7 strains generated using the split-ura4 system showed the expected localization of mECitrine fluorescence on the vacuole membrane.

Table S1 lists oligos used for plasmid construction.
Table S2 lists oligos used as donors or as primers for donor amplification.
Table S3 shows the results of PCR and Sanger sequencing analysis of Ura+ transformants obtained in the temperature-sensitive (ts) mutation knock-in experiments.

File S1 is the plasmid map file in GenBank format for pDB4279.
File S2 is the plasmid map file in GenBank format for pDB4280.
File S3 is the plasmid map file in GenBank format for pDB4281.
File S4 is the plasmid map file in GenBank format for pDB4282.
File S5 is the plasmid map file in GenBank format for pDB4283.

History

Article title

A Cloning-Free Method for CRISPR/Cas9-Mediated Genome Editing in Fission Yeast

Manuscript #

G3/2018/200164R1

Article DOI

10.1534/g3.118.200164

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    G3: Genes|Genomes|Genetics

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