...of antibiotic-resistant cells; during the time required to grease, place and fill a dozen cloning rings...endocrine cell lines we most commonly work with, require extra serum to survive at low densities - adding...growing cells such as HEK and CHO do not seem to require this, while nearly all endocrine cells do. While...

...single 96-well plate for HDR that satisfy all requirements. To alleviate this bottleneck, we began building... your own purposes, or delete the originals if desired. It works as a whole or in part. For example, you... were optimized for HDR, except for TagIn. HDR requires the additional step of mutating guide sequences...

...ethical concerns of ESCs. As iPSCs can be sourced directly from a patient’s adult cells when used therapeutically... are low. Further, as iPSC generation does not require embryos, their use skirts issues surrounding the...formation, commonly referred to as reprogramming and, direct conversion from one somatic cell type to another...

...yields from entire outgrowth built from 2 µl of the assembly reaction, and from the entire assembly reaction...assemblies) translate to faster assembly workflows that require less screening to confirm the accuracy of assembly...

...3C-based sequencing requires a large number of cells (~5x107). Together, these two requirements prevent the use...original CAPTURE method The original CAPTURE method requires the creation of a stable cell line that co-expresses...

...syndrome coronavirus 2 (SARS-CoV-2), the virus may acquire mutations to escape these therapeutics. Jesse Bloom...the antibodies were able to immunoprecipitate the entire complex in three different cell lines. The Gräslund...enzyme. Compared with previous research, which required protein compartments to co-localize multiple enzymes...

...This will make it difficult to properly gate your desired cell population. By compensating between fluorescent...other corners of the dot plot, compensation is not required.  In Figure 3, we also know that on most cytometers... cytometers, FITC and APC should not require compensation between each other. We can therefore conclude...

...level. The heartbeat – a muscle contraction – is directly related to the electrical impulse of the cardiac... to measure membrane potential and can also be directed to specific types of cells, such as neuron subtypes...For in vivo imaging applications, the ability to direct your voltage sensor selectively is especially essential...

...deposited their plasmid with Addgene, hoping to inspire others to explore the phenomenon, improve the tool...Evolution of a new variant Maria and the team set up a directed evolution and screening experiment with three ...topics. He embraces open science, self-publishing directly on his website (and often posting updates on social...

...alternative approach to NHEJ is to utilize the Homology Directed Repair (HDR) to knock in corrective genes. For...to repair the double strand break via homology directed repair and you would probably need to use a two...human cells being subjected to scarless homology-directed genome editing (Maggio et al., 2014). Gonçalves...

... minor pre-recombined population that does not require the recombinase for transgene expression. Once ... of quality control is recommended but not yet required for clinical-grade materials, and is not commonly...https://doi.org/10.1089/hum.2019.277 Lecomte E, Tournaire B, Cogné B, Dupont J-B, Lindenbaum P, Martin-Fontaine...

...suppressors and upregulation of proto-oncogenes ­­­can directly inhibit the ability of a cell to undergo programmed...that a number of novel small molecules can bind directly, albeit weakly, to K-Ras and inhibit its activation...A single mouse click on a gene of interest will direct the investigator to a list of all available Addgene...

...guest blogger Melanie Fox, founder and executive director of Central Indiana Science Outreach and a Postdoc...as a student. They didn’t always know the best direction to guide me because they had no experience with...Melanie Fox is currently the founder and executive director of Central Indiana Science Outreach and a Postdoc...

...sgRNAs and Cas9 are required for any application. If you want to include a homology directed repair (HDR) template...straightforward. Likewise for step 4, as this ties directly back to your specific goal. When thinking about...

...dCas9, require Zinc-finger or TAL proteins targeting a specific genomic locus, which can require multiple...genomic region was an intensive process. One method required insertion of a recognition sequence into the genomic...

...big science,” a massive, expensive project that required collaboration from scientists around the world... undermining science are even more immediate and dire than Zimmer could have imagined. Does this detract...about CRISPR alone, and Zimmer himself has written entire books about optogenetics. Defining science and ...

...to ddPCR. Droplet digital PCR (ddPCR) does not require a standard or a reference, meaning saving on reagents...20,000 droplets. Each droplet contains the material required for a mini amplification reaction to take place...for PCR experiments.  References Furuta-Hanawa, Birei, Teruhide Yamaguchi, and Eriko Uchida. "2D droplet...

...mammalian antibiotic selection gene. However, it does require a slower transformation recovery period than amp...zeocin resistance by expressing a protein that directly binds to zeocin, preventing it from binding DNA...low rate, there is evidence that zeocin is never entirely detoxified, even when Sh ble is expressed (Trastoy...

...sense of what AdVs are about – and we hope that it inspires further use of existing vectors, as well as research...response to adenoviral vector infected cells does not require de novo viral gene expression: Implications for...1996). The adenovirus death protein (E3-11.6K) is required at very late stages of infection for efficient...

...Molecular cloning requires some method of screening colonies for the presence of an insert. Traditionally...either briefly boiling the sample before use or by directly adding a small volume of the sample to the PCR...tell you whether the insert has the correct directionality. Verifying the insert sequence with Sanger ...

...from long written proposal narratives. Now they require grant seekers to use online applications with significant...to and proof of compliance with the grantor’s requirements as well as state and federal regulations. A ... worked for an organization in a position that required some involvement in the grants process and managing...

...generating reliable results from CRISPR edit data requires the help of robust software tools. As a consequence...of the edited sample. You can also download the entire analysis as a ZIP file by clicking the Download...the inferred distribution of indel sizes in the entire edited population of genomes. Hovering over each...

...recombinase recognizes FRT sites and initiates site directed gene recombination between those sites. FRT sites...with low concentrations of vectors expressing the required recombinase. The high concentration of FLEx vectors...PMC41457. 8. Schnütgen, Frank et al. 2003. “A Directional Strategy for Monitoring Cre-Mediated Recombination...

...exposed even one time. Doctors and researchers don’t entirely understand why some people develop the disease...one of the best ways to improve the traditionally dire prognosis for mesothelioma patients is to catch ...inhibitor) prevents the growth of new blood vessels required to sustain growing tumors. Studies have shown ...

...Addgene Executive director, Joanne Kamens, recently participated in a Reddit AMA (short for “Ask Me Anything..., work-life negotiation, and mentorship. We can direct people to the AMA in the future and link to it ...an organizer/promoter was that I didn’t have the direct link to send people until the day of the AMA. This...

... following injection into the bloodstream, but requires a high vector dose and is unable to reach all ...The Deverman group then looked to see if PHP.B required the same receptors that its parental AAV9 relied...that PHP.B, unlike its parental AAV9, does not require galactose and AAVR to transduce cells. The Wilson...

.... Cre-dependent expression of the SPH activator paired with AAV sgRNA delivery make the SPH mouse an attractive...neuroscientists, not all neuroscience applications require such optimized tools. This is elegantly demonstrated...., 2018). The pooled nature of this experiment requires fluorescence activated cell sorting (FACS) to ...

... sites. In cases like these, antibodies can be directly conjugated to agarose or magnetic beads with commercially... commercially available chemical agents. Directly conjugating the primary antibody has the additional ... particularly advantageous because they do not require centrifugation between washes which can cause loss...

...the libraries for you.  Starting from plasmids requires amplification, verification, and lentiviral production...efficiency can vary across cell types and generally requires optimization of many variables, such as seeding...Using pooled libraries can be intimidating and require a large amount of experimental work, but with this...

...on pEcgRNA plasmid as only two 24 nt oligos are required and the BsaI linearized pEcgRNA backbone can be... in large amounts and frozen for future use or directly ligated with the annealed 24 nt oligos to generate...the above systems are great for experiments that require only single chromosomal integration (ex: genome...