...formation via NHEJ, precise modification with Homology Directed Repair (HDR), CRISPR activation or repression,... edits compared to DNA transfection.  Homology directed repair (HDR) with Nanoblades CRISPR-based HDR ...To utilize HDR, a donor template containing the desired sequence flank by homology arms must be delivered...with CRISPR components. However, this sometimes requires intracellular injections into the cytoplasm or...

...be dull and painful to sit through (imagine the entire audience playing Candy Crush or reading email).... interviews. The idea is to ask questions in a directed way to bring out these interesting stories and... happens, but get things back on track if the direction of the interaction is not interesting to the audience...politely but firmly get them to stop talking and redirect the conversation. The person you cut off might...

... steel wire into a backbone structure of a protein, based on its atomic coordinates. These wire models...This post was contributed by Tim Herman, director of the MSOE center for BioMolecular Modeling and the...our guest blogger Tim Herman! TIm Herman is the director of the MSOE center for BioMolecular Modeling and...

...(Are the inserts of different sizes?), and the desired coverage for the library (How important is it that...  The most important factor to consider is your desired library coverage, since this will help decide what...determined experimentally. For larger libraries that require hundreds of nanograms of DNA, you will likely need...bacteria will be its own colony, and will not be in direct competition with other colonies containing smaller...

... a derivative of Cas9, but the ever-expanding repertoire of CRISPR has brought many other Cas effectors...part of the Class 1 CRISPR family, which means it requires multiple Cas effectors to carry out genomic editing... or just random mutations if the chromosome is repaired with all the original DNA. With Cas3, you don’...Mashimo, T. (2019). CRISPR-Cas3 induces broad and unidirectional genome editing in human cells. Nature Communications...

...Potential PIs Even if you’re applying to a school that requires its students to do rotations in different labs...to introduce yourself to a few members of your desired department during the application process. Many...However, an online meeting with members of your desired program will allow for you to have the time and...is right for you outside of just the program requirements: by talking to current students about the work...

...description. You check off all the boxes - you have the desired degree, you have mastered the applications they...person. Similar reasoning may also apply to other acquired skills.” It is nice when other people point out... of a time you’ve lead your team in a defined direction in the past. Maybe there was a time you noticed...list all the skills that your current or past job requires day-to-day, and you’ll get a great snapshot of...

...screen for IV deliverable AAVs? Typically AAVs are directly injected into the brain, but this tends to limit... (figure 4f) of the tTA inducer virus, but the desired sparsity of labeling is achieved with lose doses...-color labeling of neurons, because it doesn’t require the generation of a transgenic mouse. 2. Cell-... gene vectors for neurons, no transgenic mouse required (see figure 5 and supplemental figure 8 in Chan...

...also less accessible due to their expense and requirement for lab equipment. Plus they create plastic waste... discreet sizes (mini, midi, maxi, giga) which requires you to buy different kits for different purposes...nucleic acids are released by dipping the cellulose directly into a PCR buffer, where DNA or RNA can be amplified...This method takes less than 30 minutes and only requires incubating the columns with 1M phosphoric acid...

...content-aware suggestion dropdown list will let you jump directly to the type of information you are looking for...straightforward search is now even easier with a direct link to the plasmid page at the bottom of the dropdown... dropdown list: The search bar will suggest direct links to other types of searches including depositing...search for “Melina Fan” and you’ll be able to go directly to a page to see the plasmids deposited by our...

...students published their work identifying new genes required for stem and progenitor cell maintenance in planarians...Kimball et al., 2020). Benefits and Challenges CUREs inspire students to take ownership of their work and become...Biology Department at Keene State College in New Hampshire. He mentors undergraduates conducting NIH- and...Pellettieri, J. 2020. The exon junction complex is required for stem and progenitor cell maintenance in planarians...

...binding-dependent expression of the HIS3 gene, which is required for histidine manufacture and therefore growth...of interest, or a defined cDNA library (e.g. the entire proteome of a particular cell type, or a subset...at the same time in order to try and access the entire interactome of a set of proteins at once.  However...gain access to the nucleus. To circumvent this requirement for transmembrane proteins, a split ubiquitin...

... Chen co-founded Addgene in 2004. They worked tirelessly to get Addgene from a concept to an up and running...previous years, this event was held at Executive Director Joanne Kamen’s home- a testament to the familial...it! 8. Organize If you’re doing an event that requires people to be in groups, organize your troops prior... this saying is especially true. If your event requires a lot of set up or cleanup, ask for volunteers...

...hiPSCs usually require about 5-10 passages. Furthermore, working with Sendai virus requires stringent biosafety...to the process of collecting fibroblasts, which require a more invasive skin biopsy procedure. This becomes...PMC2917235. 11. Nakagawa, M., et al., Promotion of direct reprogramming by transformation-deficient Myc. ...

...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at the Janelia...single model. The first approach can lead to new directions, while the second approach could terminate with...think realistically about the time and resources required to feasibly accomplish the aims and clarify what... and Structural Biology, and now serves as the Director of Student and Postdoctoral Programs at the Janelia...

... and therefore may need fewer resources for the entire translation process. For example, during high growth...inappropriate locations or fail to make it through the entire transcript resulting in the production of nonfunctional...that their codons are more appropriate for the desired expression host. This is known as “codon optimization...Addgene now have full sequence data, we recommend directly analyzing gene sequences for codon optimization...

... the ideal codon for every amino acid across an entire polypeptide. Many researchers have created codon-optimization...necessary for proper protein folding (Stadler and Fire, 2011). Therefore, codon optimization may disrupt...1987) The codon adaptation index-a measure of directional synonymous codon usage bias, and its potential...doi.org/10.1016/s0165-0270(02)00362-x Stadler M, Fire A (2011) Wobble base-pairing slows in vivo translation...

...DNA strain which results in two DSBs that can be repaired by NHEJ restoring mCherry expression. Cells can...populations. To develop this method the lab was inspired by work that used an integrated GFP reporter that...fluorescent protein) upon CRISPR/Cas9 homology directed repair (HDR) (Glasser et al., 2016). For TREE ...different reporter proteins. Protein translation requires the start codon of AUG immediately downstream ...

...  Antisense RNAs encoded on the plasmid either indirectly inhibit replication through inhibiting translation...translation of replication machinery proteins, or directly inhibit replication by binding to the origin of...sequences within the origin of replication and are required for replication in plasmids that contain them ...Kamruzzaman M, Shoma S, Thomas CM, Partridge SR, Iredell JR (2017) Plasmid interference for curing antibiotic...

...skill developed during PhD training. Creativity is required to develop new projects and test novel approaches... all the funding in the world? What if you could hire more people? What would you do with unlimited time...over half of which, when implemented, impacted the entire company. It is important to note that the person...product team is an incredibly innovative group. Our entire company benefits from their policy! Perfectionism...