...are there. Make the most of everyone’s time by considering the level of meeting you need. Project Meetings...meetings tend to fill the shape of their container. Consider whether you actually need a full hour to achieve...FInd Tips on Picking a Research Project Read Other Inside Addgene Posts Resources on the Addgene Website...

...gene expression in a target cell. Practical considerations If you have high quality AAV, you can address...required for the transduced cell to become light responsive than fluorescent proteins required to get a good...interior, causing less particles to be released into suspension and lowering the number of viral particles that...

...applications such as super resolution, electron, and expansion microscopy. hfYFP is not only brilliantly bright...than most FPs. The group also tested hfYFP in expansion microscopy as well as correlative light and electron...stable FP for your next microscopy experiment, consider hfYFP and LSSmGFP.     Fig. 4: hfYFP retains...

... institutes, the Tech Transfer department is responsible for managing research use agreements like MTAs...beginning makes the sharing process smoother, more consistent, and more pleasant for everyone, which in turn...including MTAs, is perfect, but it does provide consistency and transparency in an otherwise messy process...

...space where BosLab exists today. Back then, it consisted of “wooden slab benches” set on top of filing ...companies and BosLab has grown into an organization consisting of over twenty members and volunteers. Transforming...biologist with a Ph.D. from UNC-Chapel Hill and has extensive experience targeting RNA with small molecules....

... perhaps surprising, yet important, factors to consider when seeking your dream lab. The principal investigator's...want a non-academic career path?  If you are considering this, make sure to choose an advisor that supports...where the topic of your research can make your transition easier – choose a lab that works on problems ...

...this solution wasn’t practical due to the low intensity of luminescent proteins. To make luminescent imaging...luciferase has a low quantum yield, and when transiently expressed in cells, generates much less signal...intravenously with coelenterazine, with increased sensitivity and faster imaging compared to previous publications...

... Temperature inducible promoters Temperature sensitive expression systems are typically less leaky than...Red flame plasmid pDawn contains the blue-light sensing protein YFI. When light is absent, YFI phosphorylates...? Here’s a list of some important variables to consider when choosing an inducible promoter. Experimental...

... mammals, which is thought to be due to their propensity to integrate into proto-oncogenes and disrupt...alternative virus, or other strategies may need to be considered if size constraints are an issue. Adenovirus...Adenovirus is an effective option if you need high, transient expression of a genetic cargo or have a very large...

...macromolecule density is equal to the buoyant force of the solution. Contaminants of lower densities are captured...did! From Addgene's very beginning, we had to consider digital accessibility for our website and online...

...repair template (sister chromatid or homologue) or extensive DNA synthesis. NHEJ also finishes repair of most...streamlined version of this pathway and does not consider the missing or damaged nucleotides that are common...Biochemistry and Biophysics and the Lineberger Comprehensive Cancer Center at UNC Chapel Hill.   References...

...you are spending on a conference? One thing to consider is the size of the conference. Large conferences...favor small conferences where the attendees stay onsite and eat together because scientists have time to...– even students and postdocs. There are many inexpensive/free places to order cards. There are a few good...

...publication in a scientific journal requires several considerations. First, it is important to establish the type...journal. An increasing number of funding bodies insist that the research data resulting from their funding...is a Developmental Editor for F1000Research, responsible for assessing the scientific content of manuscripts...

...red-shifted Ace-mScarlet, a fusion of a voltage-sensitive inhibitory rhodopsin (Ace2N) with the red fluorescent... Ace-mScarlet is a fusion between the voltage-sensitive inhibitory rhodopsin Ace2N and mScarlet. Ace2N...existing green-fluorescent GEVIs and blue-light-sensitive channelrhodopsins. Find the Ace-mScarlet voltage...

...using words for their field or technology in a consistent way. Just creating a lexicon was not enough. ... using the same words with the same meanings consistently, meetings will be shorter and more productive...during your postdoc that you enjoy and have a propensity for cross team communication, that’s an important...

...sequencing and qPCR methods are too costly and time intensive to process a large number of microbiome samples...bumble bee and honey bee microbiome. The toolkit consists of modular parts that are assembled with Golden...microbial population dynamics in the gut can provide insight into how the microbiome responds to infection, ...

...antibody, this approach can greatly increase the sensitivity of an assay by increasing the number of signaling...place over multiple days. Increased run time is considered one of the major drawbacks of the indirect method...interest - the secondary antibody needs careful consideration before selection. If all the primary antibodies...

...advantage of polyclonal antibodies is that they are inexpensive. Their short production time and the bulk purification...labs. Polyclonal antibodies also have a high sensitivity to the target protein due to their ability to...or fixation (IHC). Because of their increased sensitivity, polyclonal antibodies are particularly useful...

...that consistently receives around 200 new DNA samples every week. Here we will provide an inside look ...terminators. Detailed instructions on other factors to consider when designing your primers can be found here....

...negative regulation by using a promoter that is responsive to a repressor protein, the increase in expression... of repressor protein also increases thereby intensifying repression of the gene of interest and keeping...the increase in plasmid copy number during the transition to stationary phase, Segall-Shapiro et al. placed...

...organized lab saves time and money across the board. Consider alternative cloning strategies There are many ...methods, like Gibson assembly, are typically more expensive, so you’ll have to find a balance between time...either don’t scale or have limited scaling. The downside of high-throughput work is that you give up a ...

...might use it in the lab. What is a Fab Fragment? Consider the classic IgG antibody (Figure 1): it is composed...new components in your antibody tool box. As you consider which tools to use, be sure to think about what... other components of your experiment. These considerations will help you narrow down which antibody format...

... dsDNA. It cleaves ssDNA with high fidelity - sensitive to even a single mismatch in the target sequence...This makes it particularly useful in generating transient edits of signaling molecules (RNA) as opposed ...for targeting mRNA and useful as a mechanism of transient knockdown of gene expression. This system could...

...site-specific recombination events. The system consists of two components derived from the P1 bacteriophage...cyclization recombinase”), is a 38 kDa protein responsible for intra- and inter-molecular recombination ...sites are 34-base-pair long recognition sequences consisting of two 13-bp long palindromic repeats separated...

...Performance When targeting single genes, SAM consistently shows the highest levels of gene activation ...which are relatively large and are not expressed consistently throughout cells. VPR Description VPR fuses ... dCas9 to activate transcription. This complex consists of the VP64 activator used in other CRISPR activation...

...position, I just had a phone interview. I also transitioned between different lab groups without having ...that have lots of general questions about job responsibilities, performance evaluations, professional development...difficulties. Have your presentation on a flash drive and consider bringing your own laptop as a backup, if the company...

...science and collaboration. The field already has comprehensive reference pages and curated databases for scientists... (CeNGEN) The CeNGEN goal is to establish a comprehensive gene expression atlas of every C.elegans  neuron...Scientists monitor the effects of RNAi using four-dimensional contrast interference (DIC) microscopy and computer...

... this type of ELISA is extremely specific and sensitive to the target protein due to the use of multiple...concentrations of your protein of interest. The optical density (OD) - the measure of light absorbed by a solution...any experimental wells in that range of optical densities. If your standard curve does not have a linear...

... publication, Mary Dunlop’s lab generated a photosensitive Cre, Opto-Cre-Vvd, by splitting and joining.... Opto-Cre-Vvd is precise, demonstrating low sensitivity to ambient light, and versatile as it can be ...be activated by both low and high-intensity blue light. Unlike other split-Cre systems which can take up...

...faster assembly and disassembly dynamics, higher sensitivity to light, and better distribution of clusters...measure fluctuations of TagRFP-T fluorescence intensity due to interactions with photoactivatable fluorescent...By monitoring changes in TagRFP-T fluorescence intensity at different time points, the authors tracked ...

...article we will discuss general viral safety considerations and best practices for handling lab-grown viruses...replicate and continue the infection cycle once inside the host. While the virus can still enter (and ...into the host’s DNA. These viruses also have a propensity to integrate into proto-oncogenes, which can ...

...significant plasmid elements. Which differences are considered a QC issue is reviewed yearly at a best practices...populations. All of these topics require additional considerations to resolve. Sequencing and assembly issues ...assembly in Geneious can still provide additional insights to resolve the assembly and determine if there...

...fluorescent in-situ hybridization, methylation-sensitive restriction enzyme digestion, DNA adenine methyltransferase...living cells. Essentially this reporter construct consists of one substrate (the histone-derived peptide)... of these sites was controlled, the reporter consistently showed significantly different FRET levels between...

...requires a camelid animal facility which can be expensive to gain access to routinely, and it can be especially...the mRNA molecule, forming a ribosome complex consisting of the translated protein (sybody), ribosome,...constructed, they are used to produce a phage library consisting of phages that display the candidate sybodies...

...and expensive methods. While techniques such as  blue-white screening require X-gal, an expensive exogenously-added... chromoprotein, the lab determined the color intensities, maturation times, and fitness cost of expression...

...poster – now it’s time to practice your delivery. Consider your audience: Who will be attending the session... for Cancer Research Annual Meeting, you might consider including more background on how your virus of...practice: Rehearse your presentation with people inside and outside your field. Your colleagues, lab mates...

...If your poster material is nearing publication, consider uploading your work to a preprint server (and ... of the things we've mentioned above could be considered networking, but here are some more general tips...resume and prepare business cards. You might not consider being a student or postdoc a “business,” but having...

...batch analysis because of its ease-of-use and comprehensive reports. With CrispyCrunch, you get the all ...summarisation score".) For CrispyCrunch, we wanted to consider both factors, as a bench scientist would looking... our CRISPR 101 blog posts Learn about some considerations when designing gRNAs Find other CRISPR software...

...only help. Like any major life decision, the transition felt scary. While I knew two people from graduate...an industry postdoc is something that you’re considering. Like anything, there are pros and cons to postdocs... to enter academia, it is easier to make that transition from an academic setting—especially if job applications...

...result in appropriate levels of colony plating densities. Ligase fidelity and assembly efficiency Five...possibility. Recently, NEB researchers profiled the comprehensive fidelity of cohesive end ligation by this enzyme..."A single-molecule sequencing assay for the comprehensive profiling of T4 DNA ligase fidelity and bias...

... your viral supernatant. Baculovirus replicates inside insect cells, and its lifecycle ends in cell lysis...equipment. Hopefully this post has provided some insight on these differences in viral production and will...versus insect cells for viral production Consideration Mammalian cells Insect cells Viruses AAV and...

...and how will you ensure this?). You should also consider if there are any features of the library backbone...difficult to transform.    Figure 1: Factors to consider when creating a pooled library amplification protocol...BioRender.com.   The most important factor to consider is your desired library coverage, since this will...

...carry out a high-throughput genetic screen, it is expensive and time consuming to individually generate gRNAs...Most insertions can only be detected by labor-intensive processes like PCR or by evaluating visual phenotypes...PMID: 27052166. Kunzelmann, Stefan, et al. “A Comprehensive Toolbox for Genome Editing in Cultured Drosophila...

...in-person events change seemingly daily (and are inconsistent between universities!) This guide tells you ...to be incredibly useful, as it gives a more comprehensive idea of that advisor’s research interests which...program and faculty. Here are some questions to consider for your online meetings:  Is this department...

...you reach for that silica spin column, stop to consider some ways to purify DNA without a kit. DNA purification...advantages: they are convenient and provide uniform, consistent results. But they are also less accessible due... faster, cheaper, and had a similar level of sensitivity. Figure 2: Steps to purify DNA using a ...

...present even in common lab cloning strains. This intrinsic bacterial recombination pathway, referred to as...are due to incorrect primer sequences! Primers consist of two regions: the 3’ end binds template DNA for... blog posts Read our blog post on 5 factors to consider before choosing a cloning method Check out all...

...of looking for The One Method to Rule Them All, consider the following four questions to figure out which...than it is to design one demonstrating mitosis.   Consider also the amount and type of information you want...competitive while digital can be oversaturated. Transition from academic to informal writing can difficult...

...luck, and always keep going. Appendix DNAse Hypersensitive Sites and Intergenic CRISPRs Yours truly generated... In reality, cellular DNA is a complex three-dimensional globule of chromatin. Chromatin is a combination...bowtie2 -f -x HG19_GENOME --local -f -k 10 --very-sensitive-local -L 9 -N 1 -U GRNA_23MERS -S GRNA_HITS.sam...

...reporter enzyme. For multicolor staining, careful consideration must be used when selecting which antibodies...conjugates are used, but these are usually more expensive than unconjugated primary antibodies. Reagent...subclasses of 1° for multi-color staining Can be expensive Nanobodies   Recombinant; do not require ...

...So… why do some people still use amp? Carb is expensive (relative to amp), and for some applications, ...with Amp, timesaving for transformations. Con: Expensive.   Kanamycin Kanamycin, or kan, is an aminoglycoside...Like carb, spectinomycin is the more stable and expensive antibiotic compared to streptomycin. Spectinomycin...