...protein. Animals are first injected with a small peptide that represents a short, unique amino acid sequence...isolate individual hybridomas and ELISA is used to identify the hybridomas producing the most specific antibodies...antibody, meaning that the antibodies are largely identical between lots.  Disadvantages of monoclonal antibodies...antibody-encoding genes over time as the hybridoma cells divide. These changes can alter the antibody that is actually...produced by the hybridoma. Any genetic drive can be identified by sequencing the heavy and light chains. To ...

...more OptoNBs would allow for more uses both inside and outside cells. References Gil, Agnieszka A., et...exciting because using light to control signaling provides greater spatiotemporal resolution than pharmacological...domain of the Sons of Sevenless (SOS), a guanine nucleotide exchange factor that activates Ras. They expressed...more about optogenetics in Addgene's optogenetics guide ...

.... To make a point mutation, you design two oligonucleotide primers with the desired mutation, then use...3-4 phage proteins are needed and these can be provided via a separate plasmid or be inserted into the...insertions and deletions. The figure above shows guidelines for primer design; a desired insertion should...REPLACR-mutagenesis successfully incorporated a 60-nucleotide insertion; beyond this insertion size, primer...introduce mutations. Using commercially available high-fidelity polymerases such as KOD and Phusion, which are...

... in the human genome. The pair has focused considerable attention on transcription factors, starting...have good antibodies. The results look almost identical. Addgene: What advantages does your system have... also easy to run things in parallel. Addgene: Besides being fast and efficient, what other challenges... Myers: Only a small percentage of the proteins inside a cell have a commercial antibody against them.... to us to get them. Even though we’re happy to provide them, that never works on a large scale. The world...

...to $47,476 per year. This post discusses how a nationwide injunction against the FLSA is affecting universities...injunction against the FLSA updates was granted nationwide. Again, we at Future of Research believe that...higher than the level being raised to now - across a wide consensus of academic stakeholders. However, while...collecting data on the effects of the injunction nationwide and, at exactly one month following the injunction...plans, but have since reversed their position and decided to allow individual departments to go ahead with...

...time-consuming aspects of traditional cloning is the identification of clones that actually contain your insert...enzyme-mediated disruption of ccdB, leading to positive identification of desired clones.   ...Becomes a powerful... insert of interest. Hence correct clones are identified much more efficiently, as those that do not contain...ultimately perform the same function, there is some evidence that ccdB Survival™ can be more difficult to transform...E. coli do not contain the F plasmid (and are considered F-); however, there are a few popular lab strains...

...blogger Didem Goz Ayturk with edits and updates from Addgenies Karen Guerin and Susanna Stroik. Didem Goz ...administered in an organism. These features make AAV ideal for human therapies and in vivo laboratory experiments...therefore the transduced gene is episomal and does not divide and replicate in the same way the host DNA does...Elements Resources on Addgene.org Read Our AAV Guide Find AAV Vectors Browse All Viral Vector Resources...

...thousands of plasmids, using both mass production genome wide screens and the more traditional “one at a time”...time. We also handle the MTA, QC the plasmids, provide helpful resources, and, on the rare occasion things...organized lab saves time and money across the board. Consider alternative cloning strategies There are many ...either don’t scale or have limited scaling. The downside of high-throughput work is that you give up a ...engagement with and use of repositories like Addgene provides large-scale savings. How large? Using Doench’s...

...affects protein function (Bornhorst 2000).  One downside of polyhistidine tag-based purification, however...contamination, such as mammalian production systems, consider using these more selective substrates along with... containing a polyhistidine tag, you’ll need to decide whether to add the tag to the N- or C-terminus ...the opposite end.  If neither placement works, consider purifying under denaturing conditions. Unlike ...References  Resources on addgene.org Addgene Antibody Guide Ready-to-use antibodies from Addgene More Resources...

...even come [to grad school]!” He asked if she had considered other careers, which she hadn’t.  He then said... didn’t know if she liked science writing. She decided to start a blog to test it out and discovered she...communications planning.” To beef up her resume, she decided to complete a certificate in Strategic Communications...these difficult pieces: “It’s probably the best.” Besides working through the tough assignments, the skills...important for her work include: “Listening. Evaluating evidence. Critical thinking. Being able to learn things...

...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ...variant characteristics they wanted, the team identified Archon1, a novel opsin-based fluorescent voltage...researchers also showed that Archon1 is functional in a wide range of animal models such as mice, C.elegans, ...display technique using small scaffold proteins to identify novel binding proteins that bind to either the...because cPYP binds to BoPD. A similar set of newly identified light- and dark- binder plasmid tools were also...

...tumors may be the stars, there's plenty of action outside of tumor cells, and recent studies (Martins et ...microenvironments made of?  A tumor’s genesis is divided into 3 phases: initiation, progression, and metastasis...to the development and spread of cancer cells. Besides these stromal cell modifications, stromal cells...transportation, exosomes are tools of communication and provide malignancy to the microenvironment itself. They...malignancy, concluding that these cells may be considered "natural tumor suppressors." A growing interest...

...impact.  {% video_player "embed_player" overrideable=False, type='scriptV4', hide_playlist=True, viral_sharing...wonder if providing a viral vector service was a good idea. The scientific community is loving it. Almost 10,000...about this, but definitely not worried. I have confidence that from Business Development to Shipping we...

... of this is gospel, but we think it's worth consideration. Everything will go wrong Whenever an order ... barcodes and containers might not be such a bad idea, tightly coupling them to their contents might leave...easily as needed. In this case, the physical considerations of the laboratory make object composition a...for things will trip everyone up at some point, decide early on what to do in a namespace collision. It...of `dev_name` and `science_name`. You'll need to decide given the technical details of the problem you'...

...postdoc offices or professional organizations can provide referrals. There are bad coaches out there, so ...the one for the job. A professional coach will provide an initial consultation meeting or call for free...here are a number I have come across for you to consider. Feel free to add other suggestions in the comments...Scientist. Their motto is “We Turn Academics into Confident and Successful Industry Professionals”. This larger... 6 Steps to Submitting a Resume That Gets Seen  Guide Your PhD with An Individual Development Plan Learn...

...-lox Read our Genome Engineering Guide Learn more in our CRIPSR guide ...Hemizygous mice can be included in breeding strategies identical to those used for breeding heterozygous mice. ...strains that have been crossed, there are many considerations for effectively breeding your mice. Because...that express Cre but not loxP. With this two-part guide to mouse modeling now complete, you should be prepared...

... two approaches: single-cell RNA-sequencing to identify neuronal class and deep sequencing of neurons ...database specifically for C. elegans. This database provides detailed information about C. elegans 3’UTRs structures...imaging processing. The combination of these methods provides unique opportunities to develop computational ...self-excise, and covalently ligate their flanking peptides together (Wang et al., 2018).” With the split ...of CRISPR targeting vectors that each contains a guide RNA and repair template of your choosing (Schwartz...

...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ... than the more efficient single-stranded oligodeoxynucleotide donors (ssODN). So how can you use CRISPR...Resources page.  CasPhi is 70 kDa Cas protein identified exclusively in huge bacteriophage genomes. It...dual-targeting hybrid gRNA library can be used to identify genetic interactions between paralogs. The exon-deletion... viral aliquots from our plasmid collection to provide ready-to-use viral preps. Here are some of the ...

...across the university. Collective bargaining may provide a better deal for students in areas such as teaching...philosophical landscape of the scientific enterprise will provide challenges and opportunities for us all.  For example...efforts to better account for and track postdocs nationwide. On this level, FoR is currently compiling a ...further increases will be challenging, but will be aided by the development and implementation of new strategies...create tools to better understand human disease.  Outside the lab, he is interested in communicating science...

...transduction. Once you’ve harvested, it’s generally a good idea to titer your virus. Getting an accurate count of... blog and be sure to check out our viral vector guide pages to get up-to-date tips for your virus experiments...10325307. PubMed Central PMCID: PMC84922. 5. Ugai, Hideyo, et al. "Stability of a recombinant adenoviral ...immunotherapy." Acute Leukemias VIII. Springer Berlin Heidelberg, 2001. 291-306. PubMed PMID: 10634178. 8. Murnane... on the Addgene.org Check out Our Viral Vector Guide Pages Find Virus Protocols Order Virus ...