Primer pair containing BsmBI restriction enzyme cutting sites and application thereof

The invention relates to the technical field of gene engineering, and particularly discloses a primer pair containing a BsmBI restriction enzyme cutting site and application thereof, aiming at solving the problem that restriction enzyme cutting-agarose gel electrophoresis-gel recovery of a restricti...

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Bibliographic Details
Main Authors MA GUANGWEI, ZUO ZIYE, HAO XINCHONG, WANG TONGLIANG, LUO LIN, ZHAI XIAOFEI
Format Patent
LanguageChinese
English
Published 14.06.2024
Subjects
BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROCESSES USING MICROORGANISMS
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
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Summary:The invention relates to the technical field of gene engineering, and particularly discloses a primer pair containing a BsmBI restriction enzyme cutting site and application thereof, aiming at solving the problem that restriction enzyme cutting-agarose gel electrophoresis-gel recovery of a restriction enzyme cutting product is still needed in a process of constructing a targeted site knockout plasmid by a pSM-CRISPR-Puro plasmid. On the basis of a pSM-CRISPR-Puro vector, a primer pair (39bp) containing a BsmBI restriction enzyme cutting site is used for replacing a skeleton sequence (1889bp) in front of a gRNA scaffold sequence in the pSM-CRISPR-Puro vector, so that the pSM-CRISPR-Puro vector is constructed, and after restriction enzyme cutting-purification recovery of a restriction enzyme cutting product, the restriction enzyme cutting product can be used for construction of a targeted site knockout plasmid, and two operation processes of agarose gel electrophoresis and gel cutting under ultraviolet rays are
Bibliography:Application Number: CN202410407778