Summary of the Golden GATEway cloning kit.Stephan KirchmaierKatharina LustJoachim Wittbrodt
2.之前的CRISRP KO gRNA用的是Golden Gate Assembly NEBridge® Golden Gate Assembly Kit (BsmBI-v2) Golden Gate Assembly Protocol for using NEBridge® Golden Gate Assembly Kit (BsmBI-v2) (NEB #E1602) 3.现在RNAi所用的shRNA则需要用Gibson Assembly Gibson Assembly® Gibson Assembly Cloning -...
Third, we have created parts and a backbone compatible with Gateway cloning, which relies upon recombination (no mutagenesis required) and offers the advantage of large collections of destination plas- mids. The MK system includes a compatible entry plasmid (pENTR-BsaI-Tet) that supports the ...
The root-knot nematodes (RKNs) of the genusMeloidogyneare highly adapted obligate plant parasites. Their worldwide distribution and broad host range (parasitizing nearly every species of higher plant) contribute to their status as the most economically important group of plant-parasitic nematodes1. Ne...
The GB cloning strategy is especially suited for the construction of vectors incorporating Cas9 together with multiple guide RNAs in the same T-DNA. Here, we report the implementation of a GB-adapted gRNA–Cas9 toolbox for plants, which includes the domestication of gRNA/Cas9 elements, the defin...
Golden GATEwayCloningOne of the most frequently executed tasks for molecular biologists is the design and generation of complex DNA constructs. Recently, we established the Golden GATEway cloning kit for the fast and efficient generation of transgenesis vectors. This cloning kit allows the modular ...
cloning steps. We developed a set of vectors containing markers for basic cell organelles that enables the insertion of the gene of interest (GOI) by a single cloning step using the Golden Gate cloning approach and resulting in POI–GFP fusions. The set includes markers for plasma membrane, ...
2.3. Full-Length cDNA Cloning The cDNA sequences of identified target genes were generated by a microarray analysis. The full-length cDNA sequence of AhGLK1b was cloned by Gateway technology (Invitrogen, Beijing, China). Total RNA was extracted from young leaves of peanut Minhua-6 using the ...