... 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...

...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...

...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...

...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...

...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...

...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...

...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...

... 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...

... short-term protein expression observed using transient transfection approaches, generating cell lines...experiments in a stable cell line, as opposed to transiently-transfected cells, increases reproducibility,...eliminates the variation associated with repeated transient transfection. Some lentiviral vectors deliver ... in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health of the target cell...pipetting or inverting the tube. Aliquot 1 mL of cell suspension (i.e., 50,000 cells) into each well of the 6-...the virus will have higher levels of expression. Consider expanding populations transduced with a variety...expressors may take over the culture. To overcome this, consider generating monoclonal lines from the early polyclonal...

...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...

...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 ...

...capsids were packed with a GFP transgene and GFP intensity and transduction efficiency of neurons were measured...AAV.PHP.eB and AAV.PHP.S had 1.5- to 2-fold higher GFP intensity compared to the strains they were derived from...Additional Resources on the Addgene Blog Important considerations when using AAVs Learn more about using AAV ...

...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 ...

...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...

...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...