When preparing for cloning, how could you check that the restriction enzymes have produced the desired DNA fragments? When you combine a DNA insert and vector, which of these enzyme would we use? A) DNA polymerase B) DNA ligase C) Restriction enzymeģ. In the above example, which component is the vector and which is the insert?Ģ. If you get a “green check” and the message “everything checks out” then you should name your new assembly and hit the green “ Assemble” button.Ĭhoose where to save your new molecule and Voila! You are done!ġ. Hold down “Ctrl+Shift” to click on the ClaI and AlwNI sites again. Now “right-click” to open a new menu and choose “Invert Selection” to highlight this area and choose “Set from Selection” to finalize your backbone.Ĭhoose the insert in your assembly and switch to the JfyP Insert sequence. Holding down “Ctrl+Shift” (or “Cmd + Shift” for Mac) and click on both the ClaI and AlwNI restriction sites to highlight the area between both restriction sites. Work on setting your backbone first and using the pET-31b sequence. Open the “ Assembly Wizard” and choose “Digest and Ligate” as your assembly method. The fictional JfyP gene is inserted into the pET 31b plasmid using ClaI and AlwNI restriction sites.ĭigest and Ligate Cloning - Assembly Wizardįollow the instructions below to use the Benchling Assembly Wizard to plan and design cloning for your DNA constructs. We’ll look at an example of restriction cloning below. The cleavage can produce “sticky” or “blunt” ends of DNA that can be ligated to other DNA fragments that possess compatible ends. These enzymes cleave DNA sequences into fragments at or near specific recognition sites. Type II restriction enzymes are commonly used in restriction cloning workflows.
#Benchling vs snapgene how to
We’ll first demonstrate how to do traditional restriction cloning (digest-and-ligate) using Benchling. Integrate plasmid maps into lab notebooks Plan and design molecular cloning strategiesĬompare different cloning methods including restriction, Gibson and Golden Gate cloning
See previous worksheets on Benchling that introduce PCR and restriction enzyme digests.
A basic understanding of the concept of DNA cloning and familiarity with related concepts such as PCR, restriction enzymes, and DNA synthesis.
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