...Empty vectors for LIC typically employ type II restriction enzymes (e.g. BsaI), which cut at a specified...overhangs with a single enzyme, and remove the restriction sites from the final product (no "cloning scars...suggested leader sequence fused to your gene of interest, in frame with the start codon or tag sequences...sequence and sacB counter-selection marker (see our restriction digest protocol ). After the digest is complete...corresponding to the first base removed by the restriction digest to create the free 5' end of the cut vector...

...also like... Plasmid Cloning by Restriction Enzyme Digest Restriction Digest of Plasmid DNA Bacterial ...that have been generated by restriction enzyme digestion . Most restriction enzymes digest DNA asymmetrically...connecting the insert DNA (gene or fragment of interest) into a compatibly digested vector backbone. This...

..., like the miRNA bubble before it, offers an interesting view into how rapid innovation affects the research...evaluate the predictive power of each tool they are interested in by comparing their past results to the designs...

...Diagnostic Restriction Digest Verify a plasmid using restriction enzymes Diagnostic Restriction Digest Protocol...technique for molecular cloning, such as PCR- or restriction enzyme-based cloning. Purifying DNA from an Agarose...

...both antero- and retrograde labelling, synaptic restriction as well as controlled monosynaptic and polysynaptic....A., and Callaway, E.M. (2007). Monosynaptic Restriction of Transsynaptic Tracing from Single, Genetically...

...producing a final plasmid that expresses the shRNA of interest. C.1 Recommended Materials Material Vendor and...for plasmids that were successfully ligated by restriction enzyme digestion. However, once you have identified...use a miniprep kit to obtain DNA. 3. Conduct a restriction digest with EcoRI and NcoI: 1 μg miniprep DNA...select for cells that stably express your shRNA of interest. E.1 Recommended Materials Material Vendor and...cells that are stably expressing your shRNA of interest. F.1. Recommended Materials Material Vendor and...

...DNA from an Agarose Gel You may also like... Restriction Digest of Plasmid DNA Agarose Gel Electrophoresis...technique for molecular cloning, such as PCR - or restriction enzyme -based cloning. Protocol: Gel Purification...lanes between samples and between the ladder and nearest sample. To minimize the risk of DNA damage, it ...

...depending on the sample type and the protein of interest. You may need to try a variety of fixation methods...transfected with a plasmid to express the protein of interest, and a negative control sample such as cells that... that do not express the protein of interest....

...dithiothreitol, DTT), Invitrogen NP0009 PageRuler Plus Prestained protein ladder, Invitrogen 26619 Pipette tips...the hot plate to 100 °C. Thaw IgG standard and prestained protein ladder on ice. Reagent Preparation Prepare... that the chamber is sealed. Load 5 µL of the prestained protein ladder to the appropriate well. Pro-Tip...

...high-throughput next generation sequencing (NGS). Select a restriction enzyme for digest that will cut the shared plasmid...visualize that digestion on an agarose gel ( see restriction digest protocol here ). Lentiviral plasmids can...closed system of a crowded plate or culture. Try restricting growth to the bare minimum by growing at 30 ℃...

...get all your burning AAV questions answered.  Interested? You can learn more here - and remember, submissions...

...all activity on X, formerly Twitter. If you are interested in engaging with us on social media, you can ...

...complementary regions Note: If you will be including a restriction site at the 5’ end of your primer, note that ...the enzyme to cleave efficiently (e.g. GCGGCG-restriction site-your sequence). Protocol Video Watch the...

...attention to the details. Chapter 3 - Answering interesting biological questions using microscopy. Chpater...

...researchers who use VSV-G can deliver their genes of interest to a variety of cells and study their effects....

...Contributors of 2023 8 Great Depositor Research Stories Interested in contributing to the Addgene blog? We'd love...

...continuously updating this page with new and interesting content. Wish we had a video on something that...

... so we can add it to the table! And if you're interested in joining discussions about improving antibody...

... Wenning Qin, Molecular biologist with Biogen interested in applying CRISPR for mouse genome editing. ...

...Introduction 1:01-9:04 : What got Professor Cepko Interested in Working on the Eye 9:05-10:16 : Cell Targeting...