...(S) labeling of the cell population. Figure 2: Dense (c), intermediate (d), and sparse (e) labeling...References: del Valle Rodríguez A, Didiano D, Desplan C (2011) Power tools for gene expression and clonal...

...development and therapy. Nat Rev Drug Discov (2017) 16(2):89-100. DOI: 10.1038/nrd.2016.238 Additional resources...onset) may be best served by this type of targeted design. Alternatively, a disease model can be generated...

...remained active in up to 30% plasma. Figure 2: Human plasma longevity of SpCas9 vs GeoCas9. SpCas9...results. GeoCas9 RNP editing in mammalian cells Besides being able to take some heat, GeoCas9 can introduce...

...but also something about how science really works. 2)   Read science writing in magazines or newspapers...research, SciFund is really motivated more by the desire to help close the widening gap between science ...

...PMID: 30356044. PubMed Central PMCID: PMC6200774.   2. Nielsen, Alec AK, and Christopher A. Voigt. "Deep...of 173 known synthetic and natural genes.  In designing and optimizing their algorithm, they found that...

...majority of deposited materials to study SARS-CoV-2 would be fully open and available to industry requestors... enhances productive collaborations as well as provides visibility for grant-driven research being done...

... temporal activation of a transgene. Figure 2: FLEX Cre-Lox system.  WPRE The Woodchuck Hepatitis...this post, you’ve got the knowledge to be able to design a construct for the successful and specific expression...

...available to the community. We now offer AAV9 for 2 more constructs from Christopher Harvey’s lab. These...H134R) plasmids from Karl Deisseroth’s lab, which includes pAAV-hSyn Con/Fon hChR2(H134R)-EYFP that is both...

... stabilization of unknown bacteria. Figure 2: Christina Mork collecting samples at the hot spring...and workflows where CRISPR nuclease activity is desired and time or protocol steps can be saved by flexibly...

... sets Taking the time to identify and research 2 – 3 potential professions will highlight which skills...how can future regulations impact your work? What design variables need consideration? Enrich your knowledge...

...incorporation of biosensors, protein evolution lineages, 2-photon spectra, more elaborate spectrum analysis tools...or FP) is one of the most important steps in the design of an imaging experiment.  The choice is non-trivial...fusion protein, etc...).  There are many online guides – including an excellent series of posts by Joachim...information about fluorescent proteins.  It was designed with the goal of encouraging user contributions...while also maintaining a relatively strict database design (schema).  Registered users can submit new fluorescent...risk of inaccurate data submissions, so FPbase includes a versioning system that allows users to see when...for, the Table of Fluorescent Protein Properties provides a master list of all proteins in the database,...

...containing complementary polynucleotide chains. J Virol 9(2): 394-396. Lecomte E, Tournaire B, Cogné B, Dupont...our AAV preparations. We recently published a description of the VGS process and use cases in Human Gene...ligation and tagmentation preparation approaches described previously require a preliminary double stranding... the expression cassette that we want, it also includes a small amount of spurious DNA from the packaging...plasmids. We decided to use this to our advantage by designing a Python script that looks for pre-defined serotype...signature sequences in the data. Unlike previously described thermodynamics-based methods of serotype confirmation...

...interferon gamma (IFNγ) - and eliminate cancer cells. 2. Equilibrium - In a process analogous to the development... 35. Academic Press, 2015. PubMed PMID: 25818339. 2. Curiel, Tyler J., et al. "Specific recruitment of... recognition are a few methods by which tumors sidestep immune surveillance: Suppressing Function Antigen...target is never the answer to the multi-level barricades put up by cancer. As mentioned in this article...

...is the most charged, with 5 negatively charged and 2 positively charged amino acids. FLAG tag has been ... What are epitope tags? Epitope tags are short peptides introduced at the N or C terminus of a protein...purification or detection of the tagged protein. These peptides consist of as little as 8 or as many as several...need monoclonal GFP antibodies or guidance on designing a GFP tag Addgene has you covered!   Potential...Protein folding issues, steric hindrances, and destabilization can be brought about by the tag so it’s important...

...PMID: 30083608. PubMed Central PMCID: PMC6070312. 2. Stark, Jessica C., et al. "BioBits™ Bright: A fluorescent...resources to provide them to their students. My desire to create a set of low-cost, easy-to-use educational...protein – you first need the DNA sequence that encodes it. That DNA sequence then needs to be incorporated...one is as cheap as 10 cents, making activities designed with FD-CF reactions affordable to many schools...students. In our BioBits kit for example, we’ve designed FD-CF experiments that result in outputs that ...

...strategy, etc.) and episomal expression will last ~2-7 days at moderate levels. With regard to target cells...retrovirus, lentivirus, adenovirus, or AAV. Then, your desired DNA to be integrated can be cloned into the vector...packaged, when the vector is expressed. Once the design and construction steps are complete, it’s time ...weeks, depending on the virus, cell type, and/or desired event.   Figure 1: Transduction to expression...

...delivering an AAV construct expressing channelrhodopsin 2 (ChR2), a blue light sensitive cation channel, fused...blocks of polymers are heated up and pulled to a desired thickness. The fibers are drawn with microelectronics...whistles’ – including microscale light emitting diodes, thermal sensors, and microfluidic channels. These...using NeuroStack and the fibers as previously described, then measured electrophysiological potential ...

...Bacterial Cell Envelope. Cold Spring Harb Perspect Biol. 2(5) a000414 (2010). Cleveland Clinic. Gram Stain. https...phrase gram negative or positive being used to describe a species of bacteria, but what does it actually...bacteria. In a gram negative bacteria, the alcohol destroys the outer lipid membrane, and the thin inner peptidoglycan... is also useful in a laboratory setting. When designing a plasmid to be inserted into a specific bacterium...

...Without binding to the guide RNA, Cas9 cannot cut. 2. Bind to target DNA in the presence of a guide RNA...express a gRNA designed to direct Cas9 to their target sequence of choice (see tips for designing a gRNA) and... of the major CRISPR lingo and will be able to describe the functions of the various CRISPR/Cas9 components...responsible for processing foreign DNA as well as the RNA guides required for endonuclease function, when used for...Bind to a guide RNA The guide RNA, which will be described in more detail below, enables Cas9 to cut a specific...DNA damage is a DSB within the target DNA ~3-4 nucleotides upstream of the PAM sequence. Cas9 Species/Variants...with the appropriate PAM sequence) to modify the desired genomic locus.   Some historical notes CRISPR ...

....        Fig. 2: Reference image (left) provided to our graphic designer upon request that the...up walls of text. Since our guides are heavily used, our graphic designer made most of the Antibody Guide... quickly." Final design (right). Immediate and immense regret of the graphic designer not shown.   ...blog, website, social media, videos, newsletters, guides, and e-books – comes into play. Our goal is to ...communication between our content team and our graphic designer to make sure things are clear, accurate, and in...

...Nano-lanterns (eNL) described in Suzuki et al. improve the brightness of previous Nano-lanterns by 2-4 fold and... has precluded widespread adoption of the technology. In 2012, Saito et al. described the first Nano-...molecular and cell biologists over the last two decades, but their use can be limited by a few caveats....substrate furimazine. In addition to the previously described applications, the enhanced signal of eNL allows...autofluorescence and a high signal-to-noise ratio. Despite the many benefits of Nano-lantern technology, there...

...peer-mentoring groups at Addgene have gone virtual!   2. Practice public speaking For almost any science career...harder to build relationships when you are more desperate or needy so you can’t wait until you imminently...

...yourself greater chances for success.  Build a team of 2 or 3 Don’t go it alone.  You’ll find that no matter... own experiences founding and running miLab to describe some key lessons I learned while becoming a health...

... and whether I would be able to carry out a yeast-2-hybrid library screen; instead I thought they would...Coordinator’ came up at Abcam. Unlike other job descriptions for marketing positions posted on recruitment...

...MagFRET: The First Genetically Encoded Fluorescent Mg(2+) Sensor. PLoS One. 2013 Dec 2;8(12):e82009. doi: ...beginnings of the field. Those engineered sensors were designed to replace synthetic calcium dyes, which had been...

...created for the Brunello library is shown in Figure 2. You can use a small PCR product because the plasmids...next-generation sequencing (NGS). For gRNA libraries, you’ll design primers to create 200-300 bp sequencing products...

...automated. Unlike gel electrophoresis, you only need 1-2 ul of sample and the run time is just a few minutes...store your plasmid DNA Learn about the different grades of plasmid DNA preps Resources on Addgene.org ...

... remember that a resume is typically no more than 2 pages long. Write your resume for the job posting ...University of British Columbia, Student Services - includes videos on general resume writing, as well as additional...

...axon terminals to the soma (retrograde tracer) (Fig. 2). Viruses as neuronal scalpels Viruses can be injected...trillion connections (synapses.)  In this post we’ll describe the AAV retrograde serotype (AAVrg) and how it... this can cause challenges for the experimental design as these vectors can exhibit spurious expression...efficiency can have a major impact on experimental design and interpretation of results. Take extra care ...References 1. Tervo, D. Gowanlock R., et al. "A designer AAV variant permits efficient retrograde access...

...account when simulating a digest as depicted in Lane 2. Controlling methylation Finally, you can control... NEB's Resource describing Dam and Dcm Methylases Discovery of DpnI Review describing dam-/dcm- E. coli...mutagenesis. During this process, incorporation of a desired mutation into your plasmid of interest by PCR generates...only the non-mutated and methylated template is destroyed leaving behind a pool of mutated plasmids which...