...isotype converted, or chimeric, antibodies may be the right solution for you. Read on to learn more. What is...is an isotype? Antibodies are composed of light chains and heavy chains made up of variable regions and...097037-0.00017-8     Morelock, M. M., Rothlein, R., Bright, S. M., Robinson, M. K., Graham, E. T., Sabo, J...Polyclonal Antibodies Antibodies 101: Selecting the Right Antibody  ...

...Bundling up for winter? The weather outside might be frightful, but browsing the 49 new preps available ...absolutely delightful. Grab a hot beverage and a blanket, then scroll on for some wintery delights!    Table...

...neuroscience with ever more efficient tools for light mediated neuronal control and we are expanding our...target photostimulation of axons and dendrites on neighboring neurons. We have also packaged hChR2(H134R) plasmids...apparent dopamine affinity, with 113050 offering high apparent affinity, and 113049 offering medium apparent...glutamate levels. iGluSnFR.A184S, a variant with higher affinity to glutamate, has been packaged into AAVrg...

...way? What ecosystem makes your heart sing with delight? From developmental biology to developing a career...example, how to use new technologies to reach the right audiences and how open access affects financial ...of scholarly publishing. And see for example the Eighth International Congress on Peer Review and Scientific...interested in the science communications world, you might consider an internship or a short program to build...

... Addgenie Julian Taylor-Parker explains why you might want to use a specialized E. coli strain for protein... to an entirely new career path, a career coach might be able to help you. Head Addgenie Joanne Kamens...genome, you need to check whether you’ve made the right one. In this post, Addgenie Melina Fan guides us...guides us through some common controls and why you might need them. Happy Reading!   Resources on the Addgene...

...This guest blog post was contributed by Seán Mac Fhearraigh, PhD, of www.phdadvice.com. I sold out, well ...Like a considerable number of researchers, I had high hopes of securing tenure in a British or Irish University...interviewed for 1 hour so having a broad enough insight was going to help me push both my scientific and...death assays. You can find Seán on twitter @Macfhearraigh.    Read more posts about Scientific Career ...

...involves measuring the absorbance/transmission of light through a liquid to determine the concentration ...liquid. Molecules absorb different wavelengths of light to different degrees and many molecules have a specific...method takes more time to set up in the lab, you might not have to do the calculation yourself as many ...disadvantages of each method and when one method might be more suited than another. Additional resources...

...to 3-loci MPE with similar high efficiencies, which is scalable for a higher number of MPE. Therapeutic...leader sequences of a given tRNA to achieve the highest efficiencies of multiplex editing at each site....Cas9-dependent off-target editing and do not cause higher Cas9-independent off-target editing in comparison...

...sequences are from AAV2 because they demonstrate high promoter activity (Earley et al. 2020), but non-...downstream AAV applications.  Tips & FAQs Due to their high GC content and hairpin structures, ITRs are prone...rapidly replicate in a growth culture and result in a high percentage of plasmid DNA with mutated ITR sequences...Next-generation sequencing and Sanger sequencing might fail to correctly read through ITR sequences. GC...

...is not just a question of being honest and straightforward, it’s an important part of being a “good scientific...work to figure them out. But, when they come to light, it’s clear that what the researchers thought was...still missing is a clear alignment of incentives. Right now, researchers are not strongly rewarded for being.... But it ought to be possible, and if we get it right, everyone in the scientific enterprise will benefit...

...amplification and verification processes we use to ensure high-quality library distribution. You can also use these...amplification 101 Library amplification seems straightforward: E. coli bacteria are transformed with the ...a typical library, Addgene usually does four to eight electroporation reactions using a total of ~400ng.... Analyzing the reads is, again, seemingly straightforward – you need to count how many times each of ...

...protocol, get a result and hope it’s close to the right answer because if it’s not, well, your grade is ...the data look like... when it looks good, that’s a high that can last for several days.” Working with CRISPR...Doench thinks that the future of CRISPR will be in high throughput functional characterization of specific...plasmids, figure out what they do and what they might be able to do and to organize it and catalogue it...

...have purified plasmid DNA, this method can be done right in your lab in less than a day. Diagnostic restriction...sequence if necessary. The example plasmid on the right has a total size of 7.3kb, including a 1.2 kb insert...recognition sequence. Most enzyme manufacturers make High Fidelity versions of the endonucleases and/or supply...you to visualize the DNA under ultraviolet (UV) light. Don’t forget to add loading buffer to your digest...

...instrumental in helping us establish consistent, high-quality protocols for lentiviral production. Bryan...Janelia Research Campus have provided valuable insight into AAV production. Finally, we are blessed to...surveyed hundreds of scientists about various ways we might expand Addgene’s services. There was an overwhelmingly...helps scientists and allows us to maintain Addgene’s high quality standards?” Under the leadership of Addgene...

...succeed in communicating. Even the most rigorous, insightful, and novel scientific research will be wasted...your past work has prepared you to contribute at a high level to their team.  When you're writing a grant...anyone else in the world to do it.  In the lab, it might mean persuasively articulating why the results you've...to be reviewed by interdisciplinary panels.  And highly successful scientists need to communicate with ...

...labeling tubes never takes longer than 5 minutes, right?) As a result, I’d always end up stressed and rushing...add in personalized steps like “check drawers for light-sensitive reagents” or “re-sterilize now.” Have ...Moving your reagents from your left side to your right as you use them can help you remember what's been... Fig. 2: Blugene moves reagents from left to right as they use them.    Prevent Interruptions If...

...Viral integration of a construct is also reliably higher than traditional transfection and random genomic...transfection method. This is because viral systems have a high infection frequency and the viral titer (the number...responses in addition to antibody response 2) very high immunogenicity 3) and long-lasting immune response...treatment and gene therapy have also been in the limelight. The FDA approved the first oncolytic viral therapy...

...BioBuilder Educational Foundation in partnership with high school teachers to boost interest and understanding...what started in Cambridge with a handful of local high school teachers receiving training modules in synthetic...fall:  The BioBuilderClub BioBuilderClub engages high school teams around the world who combine engineering...and STEM-loving students include The BioBuilder High School Apprenticeship, preparing local (Massachusetts...

...disease. CRISPR has made genomic SDM relatively straightforward, but plasmid-based SDM has lagged behind. While...well, but poor template digestion can result in a high number of background colonies carrying the unmodified...plasmid is not maintained. Recombination efficiency is high enough to negate the need for selection usually ...introduce mutations. Using commercially available high-fidelity polymerases such as KOD and Phusion, which...

...Mendenhall: The benefits are two-fold: CRISPR/Cas9 offers high efficiency of inducing double strand breaks and ...weeks. CRISPR saves you a couple of months. It’s higher throughput and faster. It’s also easy to run things...transcription factors, but there are many reasons you might want a system like this. It’s actually surprising...with this antibody problem. We thought it was the right thing to do to put these out there. It’s a simple...

...interested in following? Someone you admire for their insight and way of expressing it in meetings? The more ...advice. Your family and friends know you well and might have good ideas for who you should connect with....cautious and make sure to choose one that comes highly recommended from someone you trust. A good coach...compliment to ask someone to share their wisdom and they might just not have the time. Thank them anyway and move...

...neutral pH compartment, ­­like the cytosol, to the higher pH of the vacuole. Read on to learn more about ...enzyme. Pho8 phosphatase activity or the molecular weight shift of Pho8 from its uncleaved to cleaved form... dual color-emission biosensor named after the brightly-coloured Australian parrot. It’s comprised of ...Oligomerization Seeing Red: Simple GFP Photoconversion Light Sheet Fluorescence Microscopy Additional Resources...

...cyan, or magenta light. A gRNA with all 3 distinct hairpin sequences produces white light. Created with ...from red, blue, and green channels overlaid on a bright-field image. Image from Ma et al, 2016. Using... A gRNA with all three hairpins generates white light. All 6 gRNAs can be expressed in a single vector...

...concentrations. Higher IPTG concentration leads to higher RepA expression which results in high plasmid copy...repressor because TALEs can be programmed to bind tightly to arbitrary DNA sequences and have been shown ...

...interactions by cross-linking RNAs and proteins with UV light, followed by an antibody pulldown of a protein of...for Affinity Purification (CRAPome), to identify highly biotinylated background proteins reported by other...and identified QKI, an RNA-binding protein that’s highly expressed in neural progenitor cells. Knock-down...Response Element (IRE) RNA motif of the ferritin light chain RNA transcript. IRE motifs are bound by IRE...

... is weaker than reporter staining. Picking the right reporter for your RANBody While all three reporters... fluorescently labeled tyramide molecule into a highly reactive radical, which is then covalently linked...antigens. Spaghetti Monster Spaghetti Monsters are highly antigenic tags that have 10 HA or 10 MYC tags built... plasmids from Penn State Need help picking the right fluorescence microscopy technique to use? Check ...

...feels she was lucky to have been in the right place at the right time when she started her scicomm career...“There were jobs that I didn’t seem to have the right skills for in terms of PR types of things or communications...leverage unpaid work for paid work: “If you use it right, it could help you get your name out for stuff that...

...result in any cells, or all the colonies screened might not contain the correct plasmid. There’s a lot that... 24 bp with the template and if the sequence has high GC or AT content, increase the overlapping region...Full lawn of cells. The antibiotic in your plate might not work. Check the plates! Streak on them an E....t get any positive clone after these steps, you might want to reconsider your approach, and try a different...

... edition of “What’s New in CRISPR” - where we highlight a few of the newest CRISPR plasmids available ...cells. The “V4” version (named CasMINI) had the highest activity for gene activation  and base editing,... not modify the Cas12f1 enzyme. The result is a highly efficient Cas system with very compact genome editors...

...RNA! Double stranded? Well, that’s just for DNA – right? RNA performs almost all of its biological functions...replication process. dsDNA viruses yield dsRNA through a slightly different mechanism thought to be associated with...always around. R-loops are particularly enriched in highly transcribed regions and repetitive regions, such...RNA:DNA hybrids levels are allowed to reach very high or low levels, possibly due to dysregulation of ...

...plasmid verification, and using NCBI BLAST. Tips for high-throughput plasmid sequencing If you’re like us,...Sanger sequencing is done from glycerol stocks (overnight cultures with 15% glycerol), as opposed to purified...handling of large numbers of samples. When we grow overnight cultures for glycerol stock production and storage...can make your life much easier by selecting the right primer. This often starts by determining what features...

...systems Gamma-retroviral vectors have a relatively high transduction efficiency, but in contrast to lentiviral...small-molecule libraries and can be combined with high-throughput sequencing to perform multiplexed analysis... cassette. It can be used to produce a ~10-fold higher titer of lentiviral particles than the original...lentivirus provides significant genetic power and allows high throughput genetic modulation studies in mammalian...

...initiation sites of at least two genes with high expression and high cell-type specificity.  Strategy #3 (ProC...promoter designs, promoters from strategy #4 had the highest rate of targeted expression. Surprisingly, <1% ...tracing experiments: While synthetic promoters with high rates of labeling are ideal of gene therapy applications...

...Maya Kostman for the IGI.     Here are some highlights of the plasmids shared over the last year and... more tools, head over to this blog post that highlights five different methods to use CRISPR to detect...

...If you’ve ever been looking for just the right CRISPR vectors on Addgene and found instead ones that ...Doench reports even the greenest members of his lab, high school interns, were able to use Fragmid to design...making it easier to get the vectors that are just right for your experiment. As always, we recommend checking...components by clicking the blue text in the upper right-hand corner.      Step 3: Reviewing the ...

...location bind to Cas9 with canonical sgRNA structure (right). Created with BioRender.com. What to edit The...3 or more mismatched bases can often evade MMR (right). Created with BioRender.com. For specific prime...prime editors bound to sgRNAs only drive nicking (right). Start with nicking sgRNAs ~50 bp from the prime...2024). Figure 9: epegRNAs (left) and PE7 (right) offer two different strategies for protecting pegRNAs...

...genome is made of protein-coding genes, and you might be wondering what the rest of our genome could possibly...non-coding RNAs! You probably didn’t see that coming, right? Non-coding RNAs (ncRNAs) are transcribed but do...versatile option. It can be used to target lncRNAs with high specificity, particularly in applications requiring...Hazan J, Bester AC. CRISPR-Based Approaches for the High-Throughput Characterization of Long Non-Coding RNAs...

...just for lab specimens — there is evidence that lightning can also promote gene transfer in soil bacteria...microbial offense One way to triumph over your neighbors is to eliminate the competition, and Penicillium...specialized degradative capabilities are often tightly regulated to be active only in the presence of ...Simonet, P. (2001). Laboratory-scale evidence for lightning-mediated gene transfer in soil. Applied and Environmental...

...acknowledgements sections. Some other additions you might wish to include are a references section, contact...navigate the poster from column to column left-to-right, which is the natural way that people expect to ...illustrations above) so that the color fades to a lighter or darker version of itself from the top of the...sure that the color of your font is compatible. Use high-contrast between these colors so that your text ...

...NGG) in both distribution and GC-content, which might improve editing in AT-rich genomes. Additionally...editing efficiency for the Cpf1 orthologs was only slightly lower than that of the widely used SpCas9 and ...that AsCpf1’s off-target rate is similar to that of high fidelity Cas9s eSpCas9 and SpCas9-HF1. Both AsCpf1... cleaves relatively far away from the PAM, NHEJ might retain the PAM site. Therefore, if HDR does not ...