...yellow derivatives) A206K Interferes with dimer interface (monomeric derivatives) K26R, Q80R, N146H, H231L...biosensors has been designed to detect a variety of intracellular conditions, including ion (such as Ca2+) concentrations...available. Others: We’ve really just scratched the surface of the potential applications for GFP. It has also...strand break repair, and to label pathogenic intracellular microbes to visualize host/pathogen interactions...

...express my concerns and request a replacement. I got my replacement in the end and the issue was resolved...less suitable for troubleshooting or issuing replacement products. In order to find good and helpful answers... in you – a technical support role is the right place to live it out, but ideally without asking too many...

...progress. You will also probably have a lot more face time with your bench mentor than your scientific...lot of details, especially when you are in a new place, learning something new. So how can you remember... walking through the protocol, by going to each space in order to double-check you remember where the ...Resources Resources on the Addgene blog Trading Spaces – Visiting Scientist Edition Mentoring for Scientists...

...target is present immediately upstream of a Protospacer Adjacent Motif (PAM), the sequence NGG.  If these... within the editing window (bases 4-8 of the protospacer). While this is a small window, this system can...system.  Fig. 1: Cytidine deamination takes place on the free strand of DNA and converts a C to U ...

...recombination; SSA, single strand annealing) took place and reconstituted the EGFP expression cassette. ...fragment containing the sgRNA target sequence can be placed in multi-cloning site (MCS) between EGFP fragments...extra precautions needed when dealing with RNA (e.g. face mask, and a dust free and specially cleaned work...mice that may have no mutation, costing money and space. As a rule, screen all of your pups unless you have...

...), the inner membrane space of mitochondria and chloroplasts, the extracellular milieu, and gram negative...should be taken into account when deciding where to place an FP within a fusion construct [1,2], i.e. positioning...mitochondria, lysosomes or they are secreted into the extracellular milieu. Many of the resident proteins within...

...is by no means exhaustive, but should be a good place to start when trying to pick your perfect promoter...promoter down regulation due to methylation or deacetylation. Ubc General expression mRNA Mammalian promoter...calmodulin. GAL1, 10 General expression mRNA Yeast adjacent, divergently transcribed promoters Inducible with...orientation to your gene Although this list is a great place to start, the tables above do not delve into the...

...The source of those conflicts can frequently be traced back to one of a few trouble spots:  Communication...here is pretty intuitive: You need to be willing to face conflict head-on. That doesn’t mean you have to ...and compromising. Though there may be a time and place for avoiding, competing, or accommodating, if you...knowledge to resolve conflicts. Conflict in the workplace is inevitable, but having who can help resolve...

...consuming and difficult. Finding volunteers who had space to shoot was another hurdle. As we slowly started...reopened. But there were still many restrictions in place, most importantly, masks.  Besides the muffled voices...strict safety protocols to keep Addgenies safe. Space was at a premium, so I had to figure out a solution... we were finally refining the process down to a place I could be happy with it. Cycle 5 - The Future! ...

...double-strand break (DSB). DSBs can be caused by intracellular factors such as nucleases and reactive oxygen...for DNA repair synthesis. The ssDNA strand then displaces one strand of the homologous DNA donor and can...formation of a DNA structure referred to as the displacement loop (D loop). The recombination intermediates...synthesized portion of the invasive strand is displaced from the template and returned to the processed...

...precision editing of the dystrophin gene would take place, with minimal risk of off-target editing. In order...sites. Each paper characterizes additional, unique facets of CRISPR DMD therapy. Long et al. show that AAV-CRISPR...the desired phenotype. Precision genome editing faces the challenges above and more. One chief challenge...approval. Even with the challenges precision editing faces, it’s encouraging to see such progress in the more...

...finding yourself having issues, you’re in the right place! In this post, we’ll talk about western blot optimization... visible to the naked eye.   If you see a smiley face in your gel, or if you see smeared bands, your voltage...voltage is likely too high. If you’re seeing a smiley face at an appropriate voltage (usually 10–15 V/cm of...bleed!), cut off the upper left-hand corner, or place a double ladder on the lefthand side. Whatever you...dry the membrane, wet it with 20% methanol, and place over a light box to visualize proteins after transfer...

... site and adjacent PAM. Your knock-in design may naturally remove the gRNA sequence or place it far from...100 bp homology on each side. For optimal repair, place the loxP sites precisely at the cut site for each...the oviductal lumen, tweezer-type electrodes are placed to mediate electroporation. Ohtsuka et al. compared...

...members of your field, and present your research. Face to face meetings can make favorable impressions on potential...Medical Institute. Today's postdoctoral fellows (PFs) face a number of challenges ranging from long periods...manuscripts. 4. Balance focus and exploration Researchers face a fundamental conflict. Science is about exploration...

...reaching the detector. The pinhole is specifically placed to only allow light from the focal plane to reach...coverslip at an angle such that when it reaches the interface between the glass coverslip and a sample in aqueous...within a few hundred nanometers of the glass/water interface. Super resolution microscopy Super Resolution ...sample thin (< 15 μm) or thick? These questions will place each sample into one of four categories that are...

...plasmids and your viral packaging service in the same place, making it easier to go from selection to viral ...to order at your convenience. After an order is placed, we’ll facilitate the necessary MTA, then start...your plasmid of interest in your desired serotype, place your order when ready, and we’ll take care of the...

...Can you imagine how much space a million plasmids would fill? Two weeks ago, Addgene edged passed our... hard to picture one million plasmids. How much space would one million bacterial stabs take up? If 81...upcoming months, we will be moving into a bigger space so that we can make an even bigger impact - we are...

...depends on a surface glycoprotein – the only viral protein present on the virion surface – making it critical... system in a novel way, by shielding host cell surface proteins. Researchers at the University of Virginia...

...factors – to specific sequences of DNA has become commonplace. This ability has opened up a world of possibilities...on a version that she hopes will change the intracellular localization of RNA. “You can think of changing...tips: First, she recommends using standard BsaI in place of the high fidelity version (despite what the original...

...transfers. When Addgene entered the plasmid sharing space, we heard over and over again that the legal documents...experiments with confidence. Benefits of the UBMTA: Peace of mind Addgene has adopted the terms of the Uniform... Recipients: Streamlined Processing: When you place an order, Addgene generates the MTA and emails it...