... can be a lot of work — and what if you want to detect more than one protein in your sample? Or what if...! Rather than start over, you can remove the detection antibodies from your membrane, allowing you to...stringent stripping methods. If you are planning to detect multiple proteins through several rounds of stripping...the strength of bands from different rounds of detection. Finally, stripping and reprobing works best with...blots that use chemiluminescent or fluorescent detection, like horseradish peroxidase (HRP) or Alexa Fluor...Fluor dyes, respectively. Chromogenic detection produces colored precipitates that permanently stain the...it. Ideally, dry your membrane once before any detection steps to avoid retaining residual antibodies. ...

...chromosomes. These long range DNA interactions are often detected with chromosome conformation capture (3C) based... of the drawbacks of 3C methods, but could only detect chromatin interactions at one location in the genome...CAPTURE 2.0, an updated version of CAPTURE that detects chromatin interactions at hundreds of loci at once... The chromatin “picture” is then developed by detecting these chimeric DNA fragments by PCR or next-generation...Cross-links are reversed so that DNA fragments can be detected by PCR or deep sequencing. A drawback of the...of CAPTURE 2.0 CAPTURE 2.0’s increased rate of detecting chromatin interactions opens the door for answering...proof-of-concept experiment to show that CAPTURE 2.0 could detect multiple chromatin interactions at once. Chromatin...

...the detection of RNA, dsDNA, and now ssDNA. By comparing the ability of Cas12 and Cas14 to detect SNPs...adaptive immune systems that provide bacteria protection against infectious agents. Several enzymes behind...fantastic add-on to a diagnostic toolkit, called DETECTR, which has been created by the authors. This kit...kit uses Cas12 and Cas13 to quickly detect the presence of infectious organisms (ex: SARS-CoV-2) and genetic...SNPs using the DETECTR kit, the authors showed that improved specificity of Cas14 enables high-fidelity ...high-fidelity SNP genotyping. The development of  Cas14-DETECTR will soon enable the quick and efficient diagnoses...resources on the Addgene Blog Learn more about DETECTR Browse all of the CRISPR blog posts Download our...

...implications of using these technologies, but also the rapidity of the technologies’ adoption by the scientific...amongst the Nobel prize winners for developing the technology. Though these case studies are “old,” they lead...significantly over the last decade. He describes the detection of gravitational waves--a story I was much less...around the world. The paper describing the waves’ detection had over 1,000 authors. The chapter on deep learning...learning addresses the dilemma of creating a technology that could eventually replace scientists themselves...Addgene’s role in the rapid dissemination of these technologies, proposing that “Addgene, with its easy, fast...CRISPR-Cas plasmids, has been a key driver of the technology.” As an Addgene vet of 12 years, I’m very familiar...

... into plasmid DNA. In high throughput cloning techniques, although the specific sequences of the genetic...proteins and the availability of powerful imaging technologies, the combination of multiple fluorescent protein...-throughput cloning. MXS Chaining is one such technique and has been used to produce complex fluorescent...constructs. These fluorescent reporters can be used to detect structure and protein localization, as well as ...show robust labeling, with strong expression and detection of each fluorophore at the respective subcellular...that the separate modules combined using these techniques are free of specific restriction enzyme recognition...High Throughput Cloning Strategies Method Technology Pros Cons References Golden Gate Type II restriction...

... to) purification, detection, solubilization, localization, or protease protection. Thus far Plasmids ...proteins, which are sometimes used as tags for detection; however, those are just one (admittedly large...overexpress and purify proteins can face any number of technical challenges depending on their protein of interest...one-step purification with glutathione, can also protect against proteolysis. That’s one form of instability...it’s a good one), then your first step towards detecting, immunoprecipitating, or purifying that protein...Stability Good for purification with glutathione; protects against proteolysis, but may reduce solubililty... improves solubility; good for antibody-based detection SUMO ~100 amino acid protein  12 Stability At...

...thanks to these technologies but many questions remain. The use of other technologies such as optogenetics... cellular movements in the immune system. The techniques used here could be adapted to studying other ...labeled cells? 1. Epifluorescence microscopy This technique uses a fluorescence microscope where the light...excitation microscopy is a fluorescence imaging technique that allows imaging of living tissue up to about...from this volume. The consequence is that you can detect signal in thick specimens. See the video below ...monitor cellular movements in real time. This technique was first Introduced to immunology by three papers...papers in the journal Science in 2002. Now, this technique and the in vivo live-cell imaging (intravital ...

...not to say that the technique doesn’t have its limitations or that other technologies need to be discarded...relatively cheap (compared to many other technologies and techniques in the sciences). But how exactly does...the history of neuroscience research we have a technology that allows us to show causality in neural circuits...interest. Despite its power and versatility, the technique is remarkably straightforward to learn and relatively...behavioral optogenetics in order to implement the technique in my lab at The Rockefeller University. In this...exactly do you want to use optogenetics for? The technique is non-trivial; you don’t want to spend months...stereotaxic surgery is a neuroscience research technique often used beyond just optogenetics. Successful...

...University of Science and Technology in Shijiazhuang, defended the technology in a news article in Nature..., NgAgo. As you might expect with any nascent technology, researchers are having trouble getting NgAgo...early successes, and to find out what delivery techniques researchers are using. The detailed results of...Mary Gearing and discusses a newer, CRISPR-based technique to generate C->T or G->A transitions in a target...target sequence in live cells. This technique uses catalytically inactive CRISPR endonuclease (dCas9) fused...in this informative post. Interestingly, this technology may make some uses of Site Directed Mutagenesis...Software for Biology Adapting Toehold Switches to Detect Zika Virus As always, if you have any questions...

...avidin-biotin technology to improve cell adhesion on nanofibrous matrices.” Journal of Nanobiotechnology 13.1 ...in molecular biology for an array of different techniques and protocols. In this post we will discuss the...interaction with avidin for molecular biology and biotechnology purposes.  Once a polypeptide is biotinylated...reporter is then added to the reaction for easy detection. The multiple biotin binding sites found on avidin...amplification during these experiments. During detection, a biotinylated reporter enzyme that is pre-incubated...amplification of the reporter signal and easier detection. This strategy is referred to as the Avidin-Biotin...Zhu, Y. “A non-radioactive method for small RNA detection by northern blotting.” Rice 7.1 (2014): 1–7. PubMed...

...in the malaria community for the better (Nat. Biotechnol., 2014). Given the timing, it shouldn’t be a ...causes malaria). I attribute their success to technical expertise, thoughtful single guide RNA (sgRNA)...yet. As a genome editing tool, the CRISPR/Cas9 technology has been surrounded by a whirlwind of research...but generally rely on less powerful algorithms to detect off-targets. The few offline tools available mostly... sgRNAcas9. Top tools for transferring CRISPR technology to a new organism: For obvious reasons you can...developed with the purpose of applying CRISPR technologies to new organisms and will help you whether you...Report your findings to help everyone else. CRISPR technology has reached into so many different communities...

...translation, but a visual technique to study translation remained elusive. Two techniques developed by Addgene...mRNAs are digested with ribonucleases, ribosomes protect the segments of mRNA to which they are bound. These...translational frequency. Although highly useful, this technique requires extensive sample processing to avoid ...-mRNA complexes. Recent work has adapted this technique to monitor local translation and determine if ... single RFP-labeled osk and Oskar protein were detected at the posterior pole. Oskar protein levels were...compartment. Despite these limitations, these techniques represent powerful tools for studying translation...Protein Plasmids Check Out Other Hot Plasmid Technologies ...

...schedule times to write.  Pomodoro technique timer  One time management technique that you can use as part of...of your writing schedule is the Pomodoro technique, which is when you spend 25 minutes focusing on a task... tomato or “Pomodoro” kitchen timer that the technique’s developer used as a university student.   ...Hemingway Editor  This is a free web-based tool for detecting the readability of your writing. It makes suggestions...suggestions Hemmngway makes (it wasn’t designed for technical writing), it’s great at flagging passive voice...jargon, this tool can help you avoid overusing technical terms when writing for a general audience. The...Simple tricks for time management: the pomodoro technique": This podcast talks more about using the Pomodoro...

...science and biotech due to its fermentation abilities and other useful traits. The “biotech yeast” You...You’re probably wondering if the “biotech yeast” is S. pombe or S. cerevisiae. Actually, it’s neither. .... The most commonly used yeast by biotech and pharmaceutical companies is K. phaffii (formerly known as... and advancements in other biomass production techniques in the following decades, Phillips made the decision...they abandoned the system. Since then, multiple biotech and pharmaceutical companies (along with some research...recombinant proteins by the pharmaceutical and biotech industries as a protein expression workhorse. This...Riley, R., et al., Comparative genomics of biotechnologically important yeasts. Proc. Natl. Acad. Sci.,...

...particularly interested in DNA sequencing tech, and the intersection of technology, software, and science. She loves... software product manager at Oxford Nanopore Technologies. If the first thing you hear when someone says...undergraduate research straight to a lab career at a biotech company, I always assumed I’d work one or two years.... Now, in my current role at Oxford Nanopore Technologies, I work with our customers in the US to learn...m always catching up in the morning! 9:00 AM--Technical implementation review. The developers discuss ...Field-based Product Manager at Oxford Nanopore Technologies for the North America region. She is particularly... for the customer and help build software and technology tools with the customer in mind. You can follow...

...to look towards genetic code expansion (GCE): techniques in which non-canonical amino acids (ncAAs, also...allow a protein to be more effectively used in a technology or as a therapeutic.  A (very) brief history ...seminal paper from Peter Schultz’s lab describing a technique that allowed for intentional incorporation of ...expansion occurred, with hundreds of new ncAAs and techniques being developed. In the mid-aughts, the first... mostly accessible to experts. That many GCE technologies have idiosyncrasies presents a significant barrier...wishing to use it as a tool. In short, while the technical skills needed to perform GCE are essentially those...Optimization Why is GCE so complicated? Well, while the techniques are straightforward, the systems – the cells ...

...transition While a PhD equips you with exceptional technical skills, transitioning into management often means...stepping into unfamiliar territory. Unlike solving technical problems in the lab, management involves a different...time, I discovered that I not only enjoyed the technical aspects of my work but also found fulfilment in...believe management is the next logical step after technical mastery. However, being a great scientist doesn...your team's—can lead to frustration. Clinging to technical tasksDelegation can feel unnatural for those accustomed...careers evolve, many professionals move between technical and managerial roles, and I've witnessed several...management and choose to focus on it, moving away from technical work. On the other hand, it’s perfectly valid ...

...Gibson Assembly Cloning and Gateway® cloning technologies, as well as several others. Each method is unique...biologists have harnessed it to evolve cloning technology. CcdB: A potent toxin... The ccdB gene, located...operon, codes for the antitoxin protein (CcdA) that protects the cell against the toxic CcdB. Cells that lose...thoroughly screen colonies for the insert. Gateway® technology (developed by InvitrogenTM) is essentially a ...check out Addgene’s Choosing a Molecular Cloning Technique page and the Addgene Blog for a variety of useful...Positive Selection of Recombinant DNA by CcdB.” Biotechniques. 1996 Aug;21(2):320-3. PubMed PMID: 8862819....Resources on Addgene.org Choose a Molecular Cloning Technique Find Plasmid Cloning Protocols Check Out All of...

...you want to read more content related to CRISPR technology, subscribe to our CRISPR blog content.   In this... dCas12f to VP64. Read the paper in Nature Biotechnology Find the Cas12f-GE plasmids Programmed genome...of target genes and cell types. Their Nature Biotechnology paper showcases an improved design of the guide...RNA motif. These engineered pegRNAs (epegRNAs) protect the RNA from degradation by cellular exonucleases...editing efficiency. Read the paper in Nature Biotechnology Find the epegRNA plasmids Enhanced prime editing...editing efficiency. The lab also optimized the architecture and amino acid sequence of the prime editor,... paired pegRNAs.  Read the paper in Nature Biotechnology Find the PRIME-del plasmids  Deletion and replacement...

... CRISPR-based cytosine base editing. Nature Biotechnology, 41(5), 663–672. https://doi.org/10.1038/s41587...activity and therapeutic application. Nature Biotechnology, 38(7), 892–900. https://doi.org/10.1038/s41587...concurrent adenine and cytosine editing. Nature Biotechnology, 38(7), 861–864. https://doi.org/10.1038/s41587...engineered Cas9-cytidine deaminase fusions. Nature Biotechnology, 35(4), 371–376. https://doi.org/10.1038/nbt...optimization and ancestral reconstruction. Nature Biotechnology, 36(9), 843–846. https://doi.org/10.1038/nbt...editors generated from TadA variants. Nature Biotechnology, 41(5), 686–697. https://doi.org/10.1038/s41587...editor with low off-target activity. Nature Biotechnology, 41(5), 673–685. https://doi.org/10.1038/s41587...