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.... One popular approach is to fuse a protein of interest to each part of a split fluorescent protein (FP...the behavior of the split FP in your system of interest. Remember that the FP fragments themselves can...

...considerations: Design your gRNA to overlap the CpG of interest. Having the CpG directly inside the gRNA sequence...but if this is not possible due to gRNA design restrictions, anything within 5 base pairs should work well...

...from Golden Gate Cloning, in which a type IIS restriction enzyme and a ligase are used to assemble multiple...described with “it’s complicated.” Applying MoClo Interested in using MoClo? You don’t have to start from ...

...noise and ensure you are measuring the events-of-interest. Sensors suitable for ER measurements include:...scientists to focus on specific cell populations of interest. Many of the calcium sensors described are also...

..., quantify, deplete, and visualize proteins of interest (Greenfield, 2014). Traditionally, they have been...similarities to host proteins. After the antibody of interest has been cloned into an expression plasmid, the...

...outskirts of Guangzhouon west of beautiful Mao Fengshan Forest Park. As a national science and technology park...surprising” small RNAs can be and posited that “we underestimate living things.” He further cautioned the distinction...

...in-situ hybridization, methylation-sensitive restriction enzyme digestion, DNA adenine methyltransferase...construct specifically to the histone protein of interest (in the figure above, H3 at K9 or K27). Joined...

... are all wired up and expressing your opsin of interest, it’s time to get to the fun stuff: the experiments...Jeanne Kreek at the Rockefeller University.  His interests include the role of learning and memory in opioid...

...student at Harvard University. Do you have a gene of interest but have run into a wall trying to study it? It...the Classical Genetics Simulator website. One interesting thing to know is that males do not recombine ...

...selected to preferentially target your cell type of interest. Viral pseudotyping is also a powerful tool that... optimize them for targeting your cell type of interest. For in vivo imaging applications, the ability...

...fusion protein containing VP2C and a protein of interest (POI). The VP2C fusion protein anchors to VP1,...into a compartment encapsulating the protein of interest (Figure 3).   Figure 3: Schematic representation...

...DNA to verify your plasmid via sequencing or restriction digest. Chances are, you’ll even have DNA left...Transformation Learn How to Perform a Diagnostic Restriction Digest Additional Resources Sigma-Aldrich Introduction...

...localization of the FP (fused to the protein of interest), thus tracking the localization of that biomolecule...at Albert Einstein College of Medicine. He is interested in everything related to gene expression, particularly...

...mini-transposon), which contains the DNA cargo of interest flanked by ~50-150 bp transposon end sequences...active in various gram-negative bacterial hosts of interest, including within mixed communities and into species...

...Scientists Making It Happen, is an exercise in restraint. The very first subheading in Chapter 1 asks “...would be complete without the inclusion of some interesting case studies, and Zimmer provides several. In...

...showing that AAV-mediated delivery of the RPE65 gene restores vision in the dog model of LCA (Lancelot, one ... again 3. Acland G. et al., 2001. Gene therapy restores vision in a canine model of childhood blindness...

...curve? A poor standard curve leads to over or underestimation of an unknown sample’s concentration. To determine...within the linear portion of the graph. It does restrict the dynamic range of your standard curve, so if...

...isolate bacteria containing your constructs of interest. But does it matter which antibiotic resistance... retained a functional copy of your plasmid of interest, preventing this issue. The genes and their antibiotics...

...biologists working with vectors are, in most cases, interested in transferring genes to mammalian cells (i.e... the viral RT protein (Faras et al., 1973). Interestingly, retroviruses are generally only “pseudodiploid...

...insert. Traditionally this has been done with restriction enzyme digest; however colony PCR can accomplish...Additional Resources on the Addgene Blog Learn to do Restriction Cloning Try SLIC cloning Learn about Gibson Assembly...

...neurological deficits observed in Rett Syndrome. Restoring the MECP2 gene by excision of the Stop cassette...precise amount of protein to each brain cell to restore physiological levels. Much research is now focused...

...scientists can make mutations throughout a gene of interest based on an organisms codon usage bias to increase...when do you want to codon optimize your gene of interest? Codon optimization has become extremely useful...

...experiments with/for them. Though not the most prestigious role in a company, Technical Support is essential... open to experienced professionals with strong interest in the technical aspects of science and the drive...

...limit the utility of candidate capsids.  Are you interested in using AAV-PHP.B vectors? See Supplementary...B capsids." bioRxiv (2019): 538421. If you’re interested in how PHP.B performed in non-human primates,...

...understandable to scientists outside your field, and interesting. For example, “there is a significant need for...Campus of the Howard Hughes Medical Institute. His interests include the quality control of secretory proteins...

... the ELN is reproducible, and only the highly restricted number of people who can access the ELN have ... the ELN arises here. Notwithstanding access restrictions based on permissions, however, in some cases...

...Let's Go! How to Verify Your Plasmid Using a Restriction Digest Analysis Videos What is a Plasmid...Electrophoresis Inoculating Liquid Bacterial Cultures Restriction Digest Analysis We also introduced guides...

... answers to such questions, Addgene is deeply interested in pursuing them. We’ve recently revamped our...can help others as much as they've helped us.  Interested in providing feedback on AAVs at Addgene? Suggest...

...envelope proteins from another virus to either restrict or broaden the host cell range (tropism). Pseudotyping..., Young JAT, Callaway EM (2007) Monosynaptic Restriction of Transsynaptic Tracing from Single, Genetically...

...every lab, there are certain hierarchies and restrictions to data access. If these are important in your...Pavlek is a Product Marketing Manager at SciNote, interested in combining scientific research with tech advances...

...could be used to routinely deliver sequences of interest to cells with efficiencies of 1-40% after G418...designing homology arms with the modification of interest, ordering them from a gene synthesis company such...

...between a pir-containing donor and the recipient of interest. Because pDS132 cannot be replicated in the recipient...Depending on the growth rate of the organism of interest, from the day primers arrive, the desired chromosomal...

...protein. SunTag has two components: 1) a protein of interest that’s tagged with 10-24 copies of the short epitope...portion of the receptors recognizes an antigen of interest while the intracellular T cell receptor portion...

...standard curve with a pure sample of your protein of interest (POI). This approach is reliable, but it’s overkill...BioRender.com. On this gel, the protein of interest band is significantly lighter in “condition 1”...

...favorite Cas enzyme, a guide RNA to your site of interest, and a donor DNA molecule containing your desired... these numbers may vary based on your locus of interest and experimental system.  Synthesizing ssODNs ...

...trainees at low wages will still be there without restructuring. Positive Points for Trainees - Taking Steps...of Research Website Additional Publications of Interest Can Be Found Here Resources at Addgene  Read...

...development of the corneal epithelium. For scientists interested in neuroscience and development, Brainbow is ...organization of the brain and other biological systems. Interested in applying Brainbow to your research? Check ...

...and repair pathways work downstream of Tet1 to restore the DNA after cytosine removal. Recently, Albert...Validated gRNAs to Target Your Genomic Locus of Interest Browse the CRISPR Collection ...

... Zinc finger nucleases (ZFNs) are artificial restriction enzymes generated by fusing a zinc finger DNA-binding...novel base editor proteins. She is particularly interested in combining the realms of art and science. Follow...

...joining for recombinant plasmid construction (restriction cloning), adaptor ligation for next generation...(LDR), probes complementary to the sequence of interest are annealed to target DNA (genomic DNA or a PCR...

...was in the lab, if you read a paper and saw an interesting construct you wished to use, you could either...Voltage Sensor  Ingaki and colleagues (8) were interested in using photoactivatable, optogenetic accuators...

... can inhibit important reactions like PCR or restriction digests. Transfection Grade DNA (high quality...than half 70% recycled materials Sustainable forestry certification Starch-based compostable bags for...

..., 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...

...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...

...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 ...

...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....

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

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