...precipitates that permanently stain the membrane, making that section of the blot unusable for reprobing... if the mild is not effective. You may choose to make your own stripping solution using a recipe like ... Assess the stripping Now it’s time to check to make sure the antibodies from your first round of detection...

...Requiring an additional plasmid for transfection can make these optimization experiments a bit more challenging...experimental volumes accordingly. While you may need to make some final adjustments, this is generally a good...additional resources. References Kumar M, Keller B, Makalou N, Sutton RE. Systematic determination of the packaging...

...vehicles for genetic information. Therefore they make excellent tools for genetic engineering. There are...vectors when used in experiments. Retroviral vectors make up a large toolbox used by researchers for (among...shRNA or miRNA. The ever expanding CRISPR field makes wide use of retroviral delivery systems, especially...

...spatiotemporal control or an “on/off switch.” This makes them great tools for providing a global overview... of the bait protein will be biotinylated. This makes it’s impossible to distinguish proteins that directly... proteins. This extended incubation period could make it difficult to detect proteins with short half-...

...it also helps you to hone your writing skills and make a name for yourself. Pick a topic that is close ...’t you record a podcast or put some effort into making a video? Use your unique talent to communicate ...Since then, I have tried many things: photography, making videos, presenting in pubs and at science festivals...

...Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J,...plasmids from this publication at Addgene. 4. Makarova KS, Wolf YI, Alkhnbashi OS, Costa F, Shah SA, ...

...meeting in person is often much more productive and makes it easier to build relationships. Nonetheless, the...responsibilities Being a leader in an organization, you need to make sure the organization speaks in “one voice” whilst...such as leadership skills can only assist you and make you more qualified for future career opportunities...

... who keep changing their minds and have trouble making decisions. In my work as a thesis coach for graduate...be reviewed, bring them to the meeting. This will make it easy for your supervisor to support you. 3) Explain...articles on our blog Read Joanne Kamen's Post on What Makes a Good Mentor Resources on the Addgene Wesbite ...

...m going to share what I’ve learned and hopefully make your decision a little easier if an industry postdoc... your goal is to enter academia, it is easier to make that transition from an academic setting—especially...company you choose. If publishing is important to you, make sure that your project will not involve proprietary...

... any features of the library backbone that would make the library more difficult to work with. For example...bacteria can grow and compete in the same container, making it easier for a smaller, recombined plasmid to ... reference protocols There are many decisions to make when developing your amplification protocol! We ...

...modulators regulate transcription and chimeric efforts make a precise edit or insertion via a chemical modifier...but it certainly suggests that Addgene depositors make CRISPR tools that fill needs in the community. The...centers around the impact of science, but our data makes it clear that it’s also a story of community, cooperation...

...capacity and stable expression of lentiviral vectors make them an attractive option. However, safety requirements...has psuedotypes. Psuedotyping is the process of making a hybrid AAV, with the genome from one serotype...Klingler, E., Kuhn, M., Farouni, R., Lagler, S., Kalamakis, G., Lischetti, U., Jabaudon, D., & Platt, R. ...

...that commercially available cloning kits can often make the process much more streamlined, because they ...the reaction in favor of the desired assembly to make the process close to 100% efficient. Gateway cloning...Multiple inserts: There are some cloning methods that make working with multiple inserts a breeze. Among them...

...School of Medicine. In the Lindberg Lab we often make cell lines that overexpress genes of interest; more...Parents of infants will have no problem with this!) Make sure your colonies are well separated. For this,...taking both early- and late-arising clones, and making sure we always maintain a backup replicates of ...

...system), and the third post will describe how you can make your own mutant flies. Find Drosophila Resources...bristly hairs. Females do not have sex combs. To make genetic crosses properly, it is important to isolate...emerging at night. An incubator with a light cycle makes for very predictable work! When flies first emerge...

...the localization of your protein of interest is to make it detectable. Visualization can be achieved using... and their potential overlap, it is essential to make sure that the selected FPs do not influence each...your next localization experiment and invite you to make any fluorescent protein constructs you create in...

...usurped old methods or complemented them in ways that make them much easier. A special class of FPs, the FP...functionality with a single gene Although it is possible to make a timer system composed of two separate FPs (one... This one protein method simplifies cloning and makes your experimental setup less cumbersome. 3. Portability...

...Knockout/Knock-in plasmids Gene targeting methods also make it possible to insert, or knock-in, any gene, tag...instead want to create a conditional knockout. To make a conditional knockout, researchers often use the...floxed alleles. Although CRISPR is very good at making knockouts, knocking in large sections of DNA can...

...Addgene is accelerating this research by helping to make new cancer resources easily accessible to the research...found across all TCGA tumor samples is represented, making the results of any such study as clinically relevant...Addgene's cancer collection pages Scientists are making progress. New resources and technologies become...

...about using non-homologous end-joining (NHEJ) to make deletions or homology-directed repair (HDR) to create...very small homology regions (5-25 bp) for repair, making it easier to construct targeting vectors. Addgene...downregulated, MMEJ represents another method for making targeted modifications. A recent publication by...