... used to integrate the tagged AID* into genomic sites of interests via CRISPR techniques. This tool-kit...activated, Cas9 cleavage can induce DSBs within minutes. New CRISPR plasmids for Borrelia burgdorferi from...are now available. These libraries encode target sites flanked by randomized nucleotides in place of a ...

... based off of p24 quantification tend to be overestimates since they can include free p24 and defective...vectors carrying fluorescent markers are used as it eliminates the need for antibody staining. Check Out Our...multiplicity of infection (MOI) to ensure only one copy integrates, selecting by antibiotic-resistance or single...

... experiments is cutting efficiency so that the greatest effect possible will be achieved. On the other... an NG PAM.   Cas PAM sequence  Additional notes SpCas9 NGG - Cas12a TTTV Great for AT-rich...coverage in the human genome. SpRY Cas9 has PAM sites covering the majority of the genome! Want to...

...co-incubate it with an “assist peptide” that facilitates endosomal escape. Combining Cas9-PAGE with lentiviral... in rodents have not been widely successful in primates. The authors used a multispecies approach to generate...endothelial-specific tropism in rodents and broad tropism in primates. Nature Communications, 14(1), 3345. https://doi.org...

... high complexity Golden Gate Assemblies. Image courtesy of New England Biolabs.   DAD Tools The three...input a sequence, SplitSet finds optimal fusion sites to split it into multiple fragments. You can define...sequences or set a narrow search window for a site—or sites—that must be used. Each of the tools works with ...

...BioID, BirA* is fused to a bait protein and it biotinylates proteins that are within a ~10 nm radius of ...the bait proteins were tagged with. It’s best to test all combinations and pick the conditions that give...interactome. Further characterization of these candidates is critical to understanding how they interact...

...career path long ago. Use your analytical mind to test and discover what career suits you best I began ... make a trophy for the winner. We promoted the contest for about two months, and we got one submission...girl about fossils. After we talked for about ten minutes, she said, “Boy, I love science,” and I realized...

... the expression system you choose in many ways dictates the best method for introducing those CRISPR components...will want to invest some time in optimizing and testing the efficiency of CRISPR delivery. There are a ... at Addgene that have been published as CRISPR testing tools: Traffic Light Reporter System: Can evaluate...

...endonuclease gene within its own cut site1. In heterozygotes, the endonuclease cuts the wild-type chromosome...Both methods are easy to implement and have been tested in yeast5. Ecological confinement involves performing...area where escaped organisms won't be able to find mates2. For example, ongoing experiments attempting to...

... learn what motivates them (keep in mind that it can be very different from what motivates you!), know...moment while reviewing the results of my Myers-Briggs test:  my undergrads were different people! More specifically...

...you’d like to go to. During the conference Take notes during talks. If you have questions during the talk...poster.” Have a notebook handy so you can jot down notes or questions from your poster viewers and trade ...whenever you have a small break), write down some notes on the card so you remember who they are and what...

... (green or blue) within a population of timer indicates the age of the pool: the higher the ratio, the...differentiation revealed by temporal cell labeling." Diabetes 58.8 (2009): 1863-1868. PubMed PMID: 19478145....programmable synthetic lineage-control network that differentiates human IPSCs into glucose-sensitive insulin-...

...Komagataella phaffii (a.k.a  Pichia pastoris) because it secretes fewer of its own proteins, making the purification... the efficacy of our antibodies through stress testing. An aliquot of our recombinant antibodies is stored...aliquots frozen immediately post-harvest. Stress-tested antibodies that are not equally effective are shipped...

...detection of the product. As an added benefit, replicates are built into the technology. One well (containing...Have a dedicated bench with a dedicated set of pipettes for ddPCR set-up. Prepare the master mix in a ...low as 1C/s. Increasing the elongation time to 2 minutes and the denaturation time to 1 minute has been ...

... is received in the lab, although this number fluctuates throughout the year (see graph below). Addgene.... Each plate has numbers and letters to give coordinates, from A1 through H12. Each plate is also barcoded...rack that contains five levels that can hold five plates on each level. This rack is located in one of five...

...to ~4-10x linearly! This expansion physically separates molecules while maintaining their relative positions...understanding of neuronal synapses, centrosomes, parasites, and more (Bertiaux, et al. 2021; Gambarotto, ...new insights into the cytoskeleton of malaria parasites including the conservation of a conoid. PLoS Biology...

...dependencies detected. Given this new information, Misek notes that in theory, if the DepMap analysis had an equal...and references References Germline variation contributes to false negatives in CRISPR-based experiments...Appelbaum, Bege Dauda, Anna Di Rienzo, Agustin Fuentes, Stephanie M. Fullerton et al. Getting genetic ...

...to spend their days microinjecting fluorescent conjugates into cells. Fortunately, over the last 20 years... cognate protein. In some cases this property eliminates the need for removing excess dye from the sample...refinements to these systems—smaller tags, brighter conjugates, higher fluorogenicity, photoactivation etc.—...

...existing sites in the original vector. Designing oligos To add NdeI, PacI, AscI and MfeI sites between ...EcoRI and SalI sites of the vector, we design a top oligo with each of the additional sites in tandem ( ... to start at 95°C for 2 minutes. Then, gradually cool to 25°C over 45 minutes. Ligation Dilute 5μL of ...tutorial we will discuss how to add new restriction sites to the MCS of an empty vector. However, the following...set of oligos containing your desired restriction sites and add them to your existing vector. It is a couple...oligo), but this would destroy the EcoRI and SalI sites in the final vector. Order the following oligos ...Place tube in 90-95°C hot block and leave for 3-5 minutes. Remove the hot block from the heat source (turn...

...role of the MECP2 gene, and highlights some of the latest breakthrough therapeutic approaches. What is Rett... the work of the consortium, two gene therapy candidates advanced into clinical development programs in... syndrome gene therapy improves survival and ameliorates behavioral phenotypes in MeCP2 null (S51.002)...