...electroporated cells re-adhered to the culture plate within 4 hr of treatment. In some primary cell lines, endogenous...articles from April 2018. If you'd ever like to write about a recent plasmid deposit please sign up here. No...

... (2017): 836-42. PubMed PMID: 28232577. Figures 2-4 were modified from this reference. 2. Regot, Sergi...Additional Resources on the Addgene Blog Learn More about Photoactivatable Fluorescent Proteins Control Protein...

...PMID: 25307932. PubMed Central PMCID: PMC4253859. 4. Hart, Traver, et al. "High-Resolution CRISPR Screens...How Barcodes Can Be Used in Lineage Tracing Read about CRISPR/Optogentics Tools Resources on Addgene.org...

... and collaborators who have succeeded at grant writing to see what tips they found useful. Step 4: Read...

...conflicts. 4) Define in advance how you would like the problem to be resolved You know more about your work... I heard about supervisors in all of the above categories. In addition, I also learned about two other...Complainers, who spend most of their time moaning about their troubles, Silent or unresponsive clams, ... students and postdoctoral fellows are hesitant about expressing their ideas, especially if their supervisors...it is interfering with your work. Avoid talking about your emotions, such as frustration or anger, because...Listen to your supervisor’s viewpoint and brainstorm about mutually beneficial solutions Your supervisor might...the problem from their viewpoint, and brainstorm about solutions that will meet both of your needs. 6) ...

...Network? Check out Our Networking for Scientists Series 4. Become a science communicator More and more fellowship...introduction to my area, a line about the research I will be doing, and a line about what I hope to discover....themselves many times and will have useful advice about what reviewers in your area are looking for. Remember...One way to add originality to your work is to collaborate with specialists in different fields that can...the field. 3. Highlight your network and your collaborators It’s a good idea to move around to different...outcomes can include poster presentations, talks about your work, or even the successful use of your preliminary...quality application. Even following this advice, only about 50% of my grant applications have been successful...

...machinery we have been able to transform 96 samples in about 4 hours with a greater than 95% success rate! Automating... work into finding robots that’ll take a little labor off our hands. Good news! There is probably a machine...96 transformations in one run compared to doing about 50 transformations in the same amount of time by...plates in just under 3 hours or one 96 well plate in about an hour. Automating vial fiIling The majority of...35 minutes compared to a lab assisstant filling about 200 vials in 45 minutes by hand. How can you start...on each tube by hand. We’ll be sure to tell you about more lab updates as we develop them. In the meantime...

... A single Nanoblade particle can be loaded with 1-4 different gRNAs. Viral envelope proteins To alter ...resources on the Addgene blog Learn about RNP delivery Learn about CRISPR delivery methods for mammalian...Nanoblades pseudotyped with a mix of VSV-G and the baboon endogenous retrovirus Rless glycoprotein (BaEVRless...have lower off-target edits. Another great thing about Nanoblades? They’re easily produced in a BSL2 lab... guide Find CRISPR plasmids and resources Learn about viral production ...

...PMID: 18479457. PubMed Central PMCID: PMC2396198. 4. Sustarsic, Marko, and Kapanidis, Achillefs N. "Taking...can be done using FRET! Read on to find out more about this amazing imaging technique and find further ...our curated list of biosensors too, as another laboratory may have already created the sensor that you ...ability of labs to answer sophisticated questions about cellular processes. Currently, FRET experiments ... Tips for using FRET in Your Experiments Learn about Fluorophore Targeting for FRET Studies FRET Based...

...PMID: 25653845. Pubmed Central PMCID: PMC4304227. 4. Revitalizing biomedical research: recommendations...purposes of labor laws, rather than as trainees in a special category, as has been the ‘labor-of-love’ philosophy...passing and upcoming implementation of the Federal Labor Standards Act (FLSA) which mandates that all workers...event of note is a ruling in August by the National Labor Relations Board to allow graduate students who work...

...Likewise for step 4, as this ties directly back to your specific goal. When thinking about step 3, however...Delivery methods for mammalian cell lines As mentioned above, the expression system you choose in many ways dictates.... The user should review the current literature about their preferred model.  If your expression system... Check out our CRISPR Featured Topic Page Learn about CRISPR Delivery Using Ribonucleoproteins (RNPs) ...

...PMID: 25936984. PubMed Central PMCID: PMC4450362. 4. Sun, J. Y., et al. "Immune responses to adeno-associated...Vectors Featured Topic Page Learn about CRISPR-AAV vectors Learn about Addgene AAV Quality Control Process...viral infections. She is particularly passionate about HIV cure research and science communication.     ...

...Biotechnology. Critical Reviews in Biotechnology, 24(4), 155–208. https://doi.org/10.1080/07388550490904288...inducible expression systems that allow tuning of metabolic pathways. Fertility (F) plasmids: microbial sharing...available in the Addgene repository. To learn more about some other common plasmid features, such as origins...Auriol, P., Vogel, T. M., & Simonet, P. (2001). Laboratory-scale evidence for lightning-mediated gene transfer...

...PMID: 25417167. PubMed Central PMCID: PMC4243060. 4. Messina, Jane P., et al. "Mapping global environmental...allowed gene circuits to be deployed out of the laboratory in a cell-free and biosafe format that was stable...circuit-based sensors to be used outside of the laboratory. The detection threshold for these RNA sequences...sequences (3 nM), however, was quite high and above what would be expected in patient samples (e.g.: blood...

...if your insert number is a bit smaller, let’s say 4 or less. The list goes on... In this post, we’ve reviewed...articles; so if you are interested in learning more about any one of them, please click on the links throughout...laying around the lab. If not, you’ll be looking at about $50-$60 per enzyme. Once you’ve got a few of these... relatively inexpensive and can be obtained for about $70, resulting in a price of a few dollars per reaction...

...This post was updated on Dec 4, 2017. At Addgene, we continually use the Basic Local Alignment Search...such as full plasmid sequences provided by the laboratories that deposit their plasmids with us or other...mismatches and gaps in the alignment. If you are curious about the differences in the blastn programs, check out...Inside Look at NGS Plasmid Quality Control Learn about our Snapgene-powered plasmid maps. Resources on...

...transcript life, and so should be handled with care (4). For this reason, viral vectors often employ other...additional helper sequences (2-3). As alluded to above, termination and polyadenylation of Polymerase II...how the Promoter Controls Gene Expression Learn about New Cloning Techniques Read all of our Plasmids ...

...Genetics 10.11 (2002): 682-688. PubMed PMID:12404098.  4. Lin, Chi-Wang, Cindy Y. Jao, and Alice Y. Ting. "... the histone protein of interest (in the figure above, H3 at K9 or K27). Joined to this by a flexible ... in Your Experiments Post Read Other Blog Posts about Fluorescent Proteins  ...

...degradation of endogenous proteins (notably, BRD2/3/4, CDK9, TRIM24, FLT3, BTK, and ALK) by linking small...degrade any protein. We are particularly excited about the dTAG system as we can use it to evaluate the...

...PMID: 29668265. PubMed Central PMCID: PMC6002863. 4. Liang, Zhen, et al. "Efficient DNA-free genome editing...scientists to make genomic alterations, bringing about a revolution in genome engineering, with new techniques...available by both the Jacob Corn and Alex Schier laboratories. Delivering Cas9-gRNA ribonucleoproteins  An...delivery of RNPs is electroporation (A in the figure above), which generates pores in the cell membrane, allowing...as lipid-mediated transfection (B in the figure above), are in the early stages of development. David ...proteins harboring receptor ligands (C in the figure above), which result in the internalization of Cas9-gRNA...show promise for in vivo studies, little is known about the immunogenicity and stability of the Cas9 protein...