...the desired chromosomal modification can be made in 5 days. Counter-selection with sacB has revolutionized... editing in Escherichia coli." Scientific reports 5 (2015): 15096. PubMed PMID: 28060411. Thomason, Lynn... a sucrase that converts sucrose into a toxic metabolite. Cells that have undergone a double crossover...confirmation of successful single crossover – after about 48 hours, the resulting colonies turn subtly blue...blog posts Find our CRISPR blog posts Learn more about Lambda Red recombineering Resources on Addgene....

...79987/ 642 670 87,400 14.0 198 Monomer  155 4.5 72 5 miRFP703 https://www.addgene.org/79988/ 674 703...for neuronal and tumour brain imaging. Nat Commun 5, 3626 (2014). Shcherbakova, D.M. & Verkhusha, V.V....in eukaryotic cells as an intermediate of heme metabolism (Shemetov et al, 2017). Modern state-of-the-art...reporters and biosensors NIR reporters can tell us about cellular events via  fluorescence changes occurring...NIR-GECOs were developed based on mIFP6. Learn more about the characteristics, performance, and applications...

...by in vivo functional screen." Scientific reports 5 (2015). BIOFAB plasmid set 3000 Series Confronted...Translational Sequence (SITS). This sequence bypasses the 5′ mRNA cap required for eukaryotic systems and instead...PUF Assembly Kit Below you’ll find information about what these kits contain, how they can be used, and...encoded sensors enable real-time observation of metabolite production." Proceedings of the National Academy...

...as scientists (5-12 years worth!) with academic scientists who can’t help us learn about the nonacademic... fine to ask about specific jobs (that is what networking is about), but be upfront about interest in ...scientists-in-training rarely get the opportunity to learn about the myriad of diverse, nonacademic careers that ...learning new things. Building new relationships is all about learning new things from other scientists doing ...pursue someone doing a job you wish you knew more about – how do you make a connection? An Informational...alienate connections. If meeting with someone to ask about a job or a particular position at their company,...that this is a good time to ask personal questions about salary, opportunities for advancement, challenges...

...Corn, for example. Learn more about the aptamer soup in this blog post. 5. Five Popular Model Organisms...mNeonGreen, mTFP1, and mWasabi by Jennifer Tsang Learn about mNeonGreen, mTFP1, and mWasabi and find plasmids...

... How to Experiment with Enhancers and Epigenomics 5 Tips for Troubleshooting Viral Transductions Single...Engineer Kris Shamloo wrote a very interesting post about how we develop our in house lab management software...

... best case, we saw that 2 out of 3 mice carried a 5 kb insertion in the ROSA locus as assessed by Southern... mouse gene with its human ortholog (greater than 5 kb), it remains to be seen whether CRISPR is as robust... Delivery Methods Learn about Mouse Work on Exercise and Depression Read About Brainbow Resources on ...success is not always guaranteed. To learn more about this process check out our Mouse Modeling blog posts...by CRISPR. The sequence started clean and then, about 100 bps into the run, it got “dirty”, with multiple...makes use of 3 components, a guide RNA (gRNA) of about 125 nt that specifies the target, the Cas9 endonuclease...Southern blot. From the efficiency discussed above extends 4 major advantages of CRISPR mouse editing when...

...create a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed.... Boncompain et al., Nat Methods 2012. Mar 11;9(5):493-8. MXS cloning platform for imaging and flow...for further chaining. The MXS-chaining kit contains 5 empty chaining vectors plus a destination vector and...half-life of both the luciferase mRNA (3’UTR AUUUA [5 consecutive]) and protein (C-terminal CL1 & PEST)....different promoters and/or allow for insertion at the 5′ or 3′ end of the sgRNA (i.e. TOP1 or TOP2 constructs...Translational Sequence (SITS). This sequence bypasses the 5′ mRNA cap required for eukaryotic systems and instead...new plasmid tool from the past year? Let us know about it in the comments or shoot us an e-mail at blog...

...favorites, all of which have blue flames. Figure 5: How to design Easi-CRISPR repair templates using ...microRNA using longer single-stranded DNA.” Sci Rep. 5 (2015):12799. DOI: 10.1038/srep12799 PubMed PMID: ... on CRISPR Mouse Models Learn about Homology Directed Repair Learn about Cas9 Ribonucleoprotein (RNP) ... a variety of species, including the standard laboratory mouse. It’s also made targeted insertions relatively...process of conventional gene targeting is lowered to about 2-3 months. But certain models remain more difficult... a mouse embryo to create chimeric mice. This laborious process takes ~8-10 months. With CRISPR (B), you...directly into a zygote and obtain a knockout mouse in about 3 months. Image courtesy of Wenning Qin. Single-stranded...

...74279). Enhanced Nano-Lanterns  Suzuki, K., et al. (5) imaging multiple proteins at the same time in live...PMID: 27240196. PubMed Central PMCID: PMC4942401. 5. Susuki, K., et al. (2016) Five colour variants of...Blog Learn more about FRET Tips for Using FRET in Your Experiments Learn more about Fluorescent Protein...heme-containing proteins in whole animals, light at or above 600 nm wavelengths should be used. Researchers often... emits light at a tissue-penetrating wavelength above 600 nm. LumiFluors Schaub, F.X., et al. (3) designed...be excited by NanoLuc® luciferase (NLuc) and set about to create an in vivo imaging reporter by linking...

...developing countries. We are proud to introduce the 5 scientists who were awarded the Addgene-Seeding Labs...The Addgene-Seeding Labs plasmid grant is a collaborative endeavor providing plasmids to researchers in... of Zimbabwe, Zimbabwe Dr. Idah Sithole-Niang (above, left) is an Full Professor in Biochemistry with... CRISPR effect observed in bacteria. Learn more about the different CRISPR applications in plants.     ...Rodrigo Arturo Rivera Solís, Associate Professor (above, far right), thrives to make affordable biosensors...detect glyphosate via H2O2, one of the reaction’s metabolites. One useful genetically encoded H2O2 probe is... to the Addgene blog for updates and future collaborations. Many thanks to all of the awardees, all who...

... F, Biette KM, DePace AH. Mol Cell. 2015 Jun 4;58(5):718-21. doi: 10.1016/j.molcel.2015.04.025. PubMed... development at the center of her lab (pictured above). DePace holds yearly planning meetings with each... their work rather than overwhelmed or confused about what’s coming next. Goal setting and career planning...and their next steps. Addgene would love to hear about the steps taken in your lab to promote personal ...

...important information in the right place on the page. 5) Apply for the job as requested in the posting Do ... your strengths in your resume and don't be shy about asking for help. The worst thing that can happen...

...the SacB gene, allowing for negative selection on 5% sucrose. Other empty vectors for protein purification...in vitro ? These, in addition to many questions about bacteria themselves, can be answered using model...overexpressing or knocking down a gene can tell you much about its function and/or alter cellular activity in a...basic biology of a bacterial gene or redirecting metabolic pathways to produce a therapeutic compound, there...once you get it right, you can learn many things about your protein’s function, structure, and interactions...molecules, light, temperature, or even intracellular metabolite levels. Different plasmids and, often times, ...pCph8 50552 pLtetO-1 Anhydrotetracycline Jeffrey Tabor Anhydrotetracycline inducible expression of red ...

..., the sacB gene is used for negative selection on 5% sucrose plates. Exchange of the sacB gene for the... 101 blog posts Learn more about Lambda Red recombineering Learn more about counter-selection Resources...levansucrase that converts sucrose into a toxic metabolite in gram-negative bacteria. Plating on sucrose...

...317–334 . https://doi.org/10.1007/s40259-017-0234-5  Büning H, Srivastava A (2019) Capsid Modifications...engineered Learn about the parts of the AAV transfer plasmid  Resources on Addgene.org Learn about AAV Find ...plasmid! Check out this blog post to learn more about the AAV transfer plasmid! References: Naso MF...

...This is the fifth in a 5 part series on Management for scientists.  We are inundated with articles and...that their teams are working well together and collaborating effectively with other teams. While they sometimes... they are as engaged as possible. To learn more about being an effective manager, read the other posts...our podcast interview with Connie Cepko to learn about her management style. Leaders look to the future...

...Dunbar DR, Mullins JJ. Nucleic Acids Res. 2007;35(5):1402-10. doi:10.1093/nar/gkl1108. Epub 2007 Feb 6...site-specific recombination in Drosophila, read on to learn about new recombinases suitable for this system. Gerald...recombinases from the Rubin lab For more information about limitations and caveats of Cre usage: Vagaries of...

...100 µL (1 mL), 25x 100 µL (2.5 mL), or 50x 100 µL (5 mL).  Once submitted, the request will be reviewed...educational resources.  We hope you’re as excited about this expansion as we are! Happy packaging! ...

...Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse Brain. PLoS ONE 8:e76310....Pseudotyped with Viral Capsids from Serotypes 1, 2, and 5 Display Differential Efficiency and Cell Tropism after...the most common concerns we get here at Addgene about this system is off-target or spurious expression...cells that don’t express Cre or Flp, what can we do about it?  Test the serotype and promoter The first step...