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...sequence starts with a 'G', you naturally have one and do not need to add an additional 'G'. Q4: Where are ...) template, you could then: Confirm your HR by doing Restriction Fragment Length Polymorphism (RFLP) ...DMSO or without 5% DMSO. My genomic PCR didn’t work, do you have any suggestions? A14: The Zhang lab recommends...you used in the Science paper (Cong et al., 2013) do you have any suggestions? A15: To date, I have never...

... single cell genomics in zebrafish. Return to top Do you have suggestions for other plasmids that should...

...Biology. 2010 Feb;2(2):a001107. PMC PMID: 20182618 . Do you have suggestions for other plasmids that should...

...many scientific questions. But these tiny creatures do tend to come with a rather astonishing ability to...

...inducer. Since induction via estradiol and progesterone do not affect each other, these systems can be used ...random variation, or noise, in reporter expression. To do so, one places reporter expression under the control... et al. Cell. 2017. PubMed PMID: 29153837. Endosomal pH sensors for STED nanoscopy Article contributed...contributed by Eric J. Perkins Listen to Endosomal pH Sensors podcast segment The first Addgene plasmid deposit... biosensors for super-resolution microscopy.” Endosomes are historically difficult to study in living ...met some of the criteria necessary for studying endosomes, the Hell lab designed their super-resolution ... meet all of them. These probes had to target endosomes, be exceptionally bright (which is necessary for...

... only problematic word. How many different things do you simply refer to as a “tube”? We took steps towards...lab or collaborators will not have the context you do, which means you’ll need to provide that context ...

...fluorescent proteins or localized fusion proteins and do not require dedicated plasmids or constructs. It ...allow for a well-controlled environment, but they do not mimic the natural situation. Measuring photostability...

...had an industry interview before. What should you do to get ready for this interview? Maybe the above ...the same boat, but there are a few things you can do to make the experience less stressful. First, you...

...Imagine an antibody. Do you immediately visualize a Y-shaped protein reminiscent of the Addgene mascot...sure to think about what pieces of the antibody you do and don’t need, and how the protein will interact...

...during your western and your results look… funky? Do you need to start a whole new blot? Never fear: membrane...directly to buffer without a re-wetting step. Important: do not dry your membrane with antibodies on it if you...

...   Viral vectors So, what do viral vectors have to do with all of these cool tools? Calcium...

...virus to mice, part of your preparation should be doing your best to be well-rested and alert the day of...procedure. Mice move much more quickly than cell cultures do, and tired researchers are accident-prone researchers...

... cell and animal models. AAVs are non-pathogenic, do not integrate into the host genome, and can be specifically...several years (Wonjo et al., 2013). Remember that AAVs do not integrate into the host genome and therefore ...

...different epitopes on the target protein, so they do not compete with each other. This antibody is also...interest that is present. Using a Standard Curve To do that calculation, you’ll need to include a standard...

...ability to target almost every genomic locus, and to do so in vivo. Catalytically dead Cas9 (dCas9) cannot...CRISPRainbow requires very short, 11-mer gRNAs, which do not induce genome editing. If used with catalytically...

... all have their benefits and drawbacks. Labs that do not routinely produce virus may want to choose more...Additional Resources on Addgene.org Learn How We Do Viral Production at Addgene Read Our Lentiviral Guide...

...fragments. When designed correctly, the recognition sites do not appear in the final construct, allowing for precise...these sites are eliminated by digestion/ligation and do not appear in the final construct. The destination...

...their current readership. Encouraging students to do science anywhere.  Chapter 4 - JEI's mission and ...

...from Addgene 1. Location, location, location! What do buying real estate and using AAVs have in common?...each serotype can transduce multiple tissues. So how do you choose the right serotype? Keep asking questions...injection. Examining serotypes 1, 8 and 9, not only do they undergo retrograde and anterograde transport... transport can be reduced by injecting a smaller dose of AAV and using a serotype with weak CNS penetrance...

...use, of course, depends on what you are trying to do – there’s no “best” tool among them. While this seems...gRNA depends an awful lot on what you are trying to do: gene knockout, a specific base edit, or modulation...importance in design – an optimized sequence will do little if it is in the wrong place, but because the...