We narrowed to 1,017 results for: PLE;

...four-base overhangs flanking each genetic part. (A) Example Level 0 assembly. BpiI recognition sites and four-base... Level 0 acceptor (bottom) for CDS1 parts. (B) Example Level 1 assembly. The choice between different ...important that these stay frozen at -80 °C, since multiple rounds of freezing and thawing will affect bacterial...plasmids with undesired ligations, so always screen multiple colonies after each transformation. This could... MoClo user, but beginners or those performing complex assemblies should screen a minimum of 8–10 colonies...

...manipulated for use in molecular biology studies. For example the E. coli biotin ligase, BirA, has been used ...corresponds with human diseases and conditions. For example, the rat is frequently used to study human cardiovascular...abilities. The axolotl has the ability to regenerate complex structures such as limbs, the spinal cord, and ...them an ideal model organism; they are relatively simple to maintain and breed and they have large, easily... and microinjections. Injections of mRNAs, for example, can be analyzed to understand human disease alleles...

...cells will be co-transduced with multiple copies of both vectors. Examples of use: Halbert et al., 2002...then following transduction, the partial genomes complement each other to restore the full-expression cassette...outperform single oversized AAVs (Hirsch et al, 2013). Examples of use: Lai et al., 2005 used a trans-splicing... the accumulation of such truncated proteins. Examples of use:  Ghosh et al., 2008 used a hybrid split...

... words, any insertion or deletion that isn’t a multiple of three will do. A popular screening method for...trace file generated in Sanger sequencing and a simple algorithm to quantify editing efficiency by insertion...have to screen to identify a knockout clone. For example, if your cell line is diploid (2N) and your out...to compare the sequencing reads from your edited sample to an untreated population. Software such as CRISPResso... Multiple Authors...

...G base editor (ABE) followed based on similar principles (Gaudelli et al., 2017). These editors did not...which could be repaired with an A→G edit on the complementary strand. But the invention of base editing doesn...chromosomal location, sequence context, and cell type. Multiple labs worked to improve its effectiveness through... a machine learning approach, the lab proposed multiple sets of Cas9 mutations that conferred new PAM ...Jiang, W., Marraffini, L. A., & Zhang, F. (2013). Multiplex genome engineering using CRISPR/Cas systems. Science...

...protein phosphorylation in an array of model systems coupled with advances in mass spectrometry suggest that...recognizes a specific ‘open’ codon and utilizes this triplet to incorporate phosphoserine A Sep-tRNA synthetase... purified from the system, researchers can run samples on SDS-PAGE gels containing Phos-Tag, a small molecule...substitution. Alternatively, you can express the completely wild type protein from the same plasmid.  Upon...reference and protocols for use of this system, please reference ‘Rinehart Lab reagents for improved expression...

...emotions and assumptions. I’m not just talking about simple things like “Which route should I take to get to... give you a fuller picture. Consider talking to people in the lab across the hall or in the same department...choosing a training lab, I strongly suggest you complete a work rotation in the lab if at all possible.... lab members alone during the interview period. People seem to be hiding their work and are nervous about...successful. Collaborative Competitive Papers have multiple authors One author plus PI on most papers Productive...

...goal of isolating single cell clones to use for multiple downstream experiments. You only successfully ...specific list of the candidate off-target sites. For example, if there are 10 putative off-target locations ...most commonly used Cas, spCas9, has gone through multiple rounds of optimization which have led to more ...confounding your results should be minimal. For example, if your goal is to generate a knock-out cell line...are performing CRISPR library screens, the same principle holds true - the higher the number of distinct...

...us about a new tool for efficient multiplex base editing and multiplex prime editing (requiring minimal...editing tool for large inserts.  DAP arrays for multiplex gene editing: Guest blogger Qichen Yuan tells ...a research story you want to share on our blog, please reach out to [email protected] - we'd love to hear...

...journey is at an end! Their name is...(drum roll, please!)... Abi! Thank you so much to everyone who submitted... name. So we reached out to you and received a plethora of options, ranging from Affie to Yncredibody....Which name best fits our new immunoglobulin friend? Please help us decide by voting for your favorite on this...Original Blugene and Aavery would like your attention, please, as they introduce their new friend…  Fig...

...antibodies, for example through the use of less toxic adjuvants; 2) generating complex antibody chips ...cells are fused with activated B cells from the spleen of mice injected with the antigen of interest. ... assessment of many protein targets in a single sample; 3) producing recombinant, partially humanized ...

...Bernstein and Boyden, 2011; Mei and Zhang, 2012). For example, blue light activates channelrhodopsins that, in...Abdallah et al., 2018; Deubner et al., 2019). For example, Taylor and colleagues used rAAV FLEx-based vectors... Han, 2016). Not so long ago, however, it was completely unclear which subpopulation of DA neurons could.... "A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit ...Walther Stoeckenius. "Rhodopsin-like protein from the purple membrane of Halobacterium halobium." Nature new...

...methods. (A) Basic structure of an antibody. (B) Examples of engineered antibodies, including chimerization...achieve whatever goal you’re aiming for. When you complete each cycle, your best-performing mutant is usually...or predicted structure of your antibody-antigen complex as a template. This structural framework then guides...specific functionality you’re looking for. For example, if you were aiming to increase an antibody’s binding...conditions. These methods are especially powerful when coupled with iterative rounds of selection and mutagenesis...

...generate 2 possible products: 1) a partially dsDNA duplex with single-stranded 3’ overhangs or 2) if the ...dsDNA was short enough, a ssDNA whose entire complementary strand was degraded. Beta: Beta protects the...created by Exo and promotes its annealing to a complementary ssDNA target in the cell.  Only Beta expression...region of interest and design an oligo that is complementary to the lagging strand. In practice, it’s easier...system once recombinant clones are generated.  A simple way to do this is to express lambda red genes from...

...Addgene, we try to make the process as smooth and simple as possible. We are the Customer Support Specialists... Placing an order on Addgene’s website is quite simple, even if you’re using a PO number. Follow these...generated through your organization in this field. Please also enter the billing address for your organization...time, your organization will have net 30 days to complete payment for your order.  Still have questions?...

...competition is fierce. However, I believe there are plenty of great jobs out there for science PhDs. The problem...organizers are building valuable skills and meeting new people by working together on the event planning and execution... execution. I am seeing many similar examples of science trainees taking a proactive role in their non-bench... Consulting. I know of many schools that have completely student/postdoc-driven biotechnology clubs such...

...visualization in vivo using ultrasound (for an example see the figure below). In addition to low costs...benefits of this technique. It is even possible to multiplex ultrasound dyes because these dyes “collapse” ...Approaches to Presenting the Microbial World’ showcased complex connections and ideas in various visualized approaches...gifs created by the Amoeba Sisters. Here’s one example of how they “bridge the jargon gap using gifs”:...

... 9. Droplet Digital PCR for AAV Quantitation by Kate Harten DeMaio We made the switch to droplet digital...Leah Schiesow If you're thinking about hosting multiple plasmids in one cell, it might be that you can't...Purification Without a Kit by Beth Kenkel To complement our RNA Extraction Without A Kit blog post, we've...

...cytometers. These fluidic machines handle biological samples and are cleaned between each use. You wouldn’t ...high-throughput and efficient way? Grenova, for example, offers automated, benchtop, high throughput washing...lab culture, but with the right incentives (for example, by grant agencies), we can shift toward a more...more sustainable one. The math on this is pretty simple: wash your tips once and you cut your tips cost ...

...feedback and input on multiple topics every time you meet.  Seek feedback in multiple formats      Ask ...and often A one-time event doesn't work to get people sharing.   Frequently ask direct questions for ... in a 5 part series on Introduction to Managing People for Scientists. Subscribe to the Addgene Blog to...