Thermo Fisher Two Vector Molecular Cloning Flow

Chitinase gene was cloned into pjet1 2 blunt cloning vector using clonejet pcr cloning kit thermo scientific usa.
Thermo fisher two vector molecular cloning flow. Navigate through the. Once the initial results are displayed they can be further refined by selecting among characteristics such as promoter inducer and antibiotic selection. In addition we also have a wide selection of products related to vectors including competent cells media plasmid purification kits and more. A second popular approach uses pcr to amplify the region of interest from the plasmid.
Plan on two plates per each ligation transformation reaction. Ligation into the positive selection vector takes only five minutes yielding more than 99 recombinant clones. Ta cloning or blunt end cloning methods can be used as described in the pcr cloning section but neither approach maintains directionality of the insert. The resulting pcr product is then cloned into the desired vector.
Learn about the foundations of recombinant dna technology. Step action 1 prepare either electrocompetent or chemically competent gi724 cells. Learn about the foundations of recombinant dna technology. 3 prepare rm medium and rmg amp plates.
Simply search by keyword or filter your results by vector type host system or cloning system. Simply search by keyword or filter your results by vector type host system or cloning system. Bacterial transformation and competent cell education. Find educational resources available for bacterial transformation covering the basics workflow considerations competent cell selection by applications and troubleshooting.
Traditional cloning relies on recombinant dna methods that begin with preparing a vector to receive an insert dna by digesting each with restriction enzymes. We offer everything from custom vector design to high throughput protein expression. The thermo scientific clonejet pcr cloning kit is a versatile advanced positive selection system for high efficiency cloning of pcr products. Nucleic acid purification and analysis support center.
All common laboratory e coli strains can be directly transformed with the ligation product. Once the initial results are displayed they can be further refined by selecting among characteristics such as promoter inducer and antibiotic selection. Simply search by keyword or filter your results by vector type host system or cloning system. Once the initial results are displayed they can be further refined by selecting among characteristics such as promoter inducer and antibiotic selection.
The digested fragments are then spliced together by an enzyme called ligase in a process known as ligation to form a new vector capable of expressing a gene of interest. To achieve directional cloning restriction sites that are present in the destination. 2 develop a cloning strategy to fuse your protein in frame with the initiation atg in the vector plex. In addition we also have a wide selection of products related to vectors including competent cells media plasmid purification kits and more.