期刊
PLANT METHODS
卷 14, 期 -, 页码 -出版社
BIOMED CENTRAL LTD
DOI: 10.1186/s13007-018-0305-8
关键词
CRISPR/Cas9; Genome editing; Tobacco; Arabidopsis thaliana; PCR; Mutant screening
资金
- National Key Research and Development Program [2016YFD0101006]
- Science and Technology project of Henan Province [172102110153]
- Project of the Tobacco Genomic Program [110201501015(JY-02)]
- National Natural Science Foundation of China [31770300]
- Program for Innovative Research Team (in Science and Technology) in University of Henan Province [18IRTSTHN023]
Background: The CRISPR/Cas9 system is being used for genome editing purposes by many research groups in multiple plant species. Traditional sequencing methods to identify homozygous mutants are time-consuming, laborious and expensive. Results: We have developed a method to screen CRISPR/Cas9-induced mutants through Mutation Sites Based Specific Primers Polymerase Chain Reaction (MSBSP-PCR) The MSBSP-PCR method was successfully used to identify homozygous/biallelic mutants in Nicotiana tabacum and Arabidopsis thaliana, and we speculate that it can be used for the identification of CRISPR/Cas9-induced mutants in other plant species. Compared to traditional sequencing methods, MSBSP-PCR is simpler, faster and cheaper. Conclusions: The MSBSP-PCR method is simple to implement and can save time and cost in the screening of CRISPR/Cas9-induced homozygous/biallelic mutants.
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