...effective method for detecting plasmid multimers. Digests themselves cannot be used to detect multimers, since...Electrophoresis (CGE) analysis Another technique that can detect multimers is capillary gel electrophoresis...upstream production. Biotechnol Adv. 2009 Jul-Aug;27(4):353-70. doi: 10.1016/j.biotechadv.2009.02.003. Epub... the plasmids, including long-read sequencing techniques, such as Oxford Nanopore sequencing or PacBio...via antibiotic selection (Crozat, 2014). How to Detect Plasmid Multimers Run an undigested plasmid on ...sequences within a plasmid, it cannot be used to detect plasmid multimers. Some companies provide NGS whole...SMRT sequencing. These longer reads are able to detect large repeat sequences within a plasmid and, therefore...

...activity mapping with STARR-seq   Although many technologies have been developed for the study of enhancer...and drug resistance. Recently, the Wu Lab at Texas Tech developed a pooled CRISPR knockout library and used...deep sequenced to identify the gRNAs that confer protection to WNV. From this screen the authors were able...assembly utilizes 5 different overlap sites, this technique also allows for easy and rapid targeting of complex... and mutant library generation in several biotechnologically and clinically relevant bacterial species...SnoopCatcher The development of recombinant DNA technology has allowed molecular biologists to fuse fragments...through Addgene. Stovicek, et al. J Ind Microbiol Biotechnol. 2015. PubMed PMID: 26376869 Jensen, et al. FEMS...

...received a Ph.D. in Biology and Biotechnology from Worcester Polytechnic Institute. What is RNAi? The loss-of-function...earned his Ph.D. in Biology and Biotechnology from Worcester Polytechnic Institute where he investigated...the inherent advantages and limitations of any technique in the context of the biological question. For...unattractive from a practical perspective. Therefore, techniques that function downstream of DNA, such as RNA ...interfering RNAs.   RNAi vs. CRISPR: Complementary techniques It is fair to say that CRISPR has revolutionized...With the meteoric rise and diversity of CRISPR technologies, one might incorrectly relegate RNAi to history...requires the presence of dsRNA. In plants, CRISPR technologies are predominantly used to disrupt function at...

...very specifically to target molecules. Using a technique called systematic evolution of ligands by exponential... used in a variety of applications, including detection of environmental pollutants, drug discovery, and...a “bad batch”); Customizable sensitivity and detection - since they frequently undergo conformational...oligonucleotides in fluorescent visualization techniques for decades, most notably for fluorescent in ...visualize RNA. Low signal-to-noise ratios with such techniques can be problematic, however. Well designed aptamers..., thereby improving traditional visualization techniques. In addition, their incredible versatility opens...Fluorescence resonance energy transfer (FRET) technology has been studied and used in assays for over ...

...disease earlier. New techniques are currently being tested in clinical trials to detect mesothelioma earlier...diagnosis, the hope is that as these new detection techniques and therapies emerge into mainstream use... asbestos. However, as the U.S. Environmental Protection Agency has noted, there is no safe level of asbestos...growth of tumors. More often than not, multiple techniques are used as part of a treatment plan, an approach...intraperitoneal chemotherapy (HIPEC). With this technique, surgery is performed to remove the mesothelioma...delivered intravenously. New treatment options: Early detection, immunotherapy, and angiogenesis inhibitors New...life expectancy of mesothelioma patients: early detection and immunotherapy.  It has long been known that...

...people find it can detect even smaller differences, and it can even be used to detect a single copy of ...which aims to increase your understanding and technical proficiency with common applications - now let... thumb, you can assume that qPCR can accurately detect expression differences of twofold or greater with...Can be multiplexed Single amplicon validation technique Hydrolysis Probes (Taqman) Low; can be reduced...Biassoni, 2014). At the bench As a highly sensitive technique, qPCR needs a steady pair of hands to produce ...of triplicates to understand variation due to technical error.  Plate readouts can be affected by plate...differences may be a concern in your reaction, analysis techniques that use raw data corrections are a better choice...

...proteins may not be detected at all, while overloaded proteins may saturate the detector. In between these...extremes lies the linear detection range. Figure 4: The linear detection range of the assay is between...loading control points are outside of the linear detection range. As a result, they have almost the same ...computational quantification. Outside the linear detection range, band intensity measurements become extremely...are often called “housekeeping proteins” and are detected with an antibodies, just like the POI. This approach...means that at sample concentrations appropriate to detect a less-abundant POI, the loading control will almost...error, it’s important to determine the linear detection range of all proteins that will be measured in...

...contaminants and to subsequently develop technologies to protect fresh water repositories in Mexico. At ...insecticidal proteins to the development of renewable biotechnology products and biosensors for herbicides as well...a bacterial expression system for renewable biotechnology products Dr. Idah Sithole-Niang, University ... Lecturer and a Plant Molecular Biology and Biotechnology expert. In order to preserve Cassava yields ...Robertson aims to make use of the CRISPR/Cas9 technology to engineer plants that are able to destroy DNA...Associate Professor (Right), focuses on molecular biotechnology for improving expression of nutritionally and...Bao Ton (center) is interested in applying gene technology to bacteria and plants. Her research involves...

... a revolution in genome engineering, with new techniques rapidly being developed. Performing a CRISPR ...potential for off-target effects. Cas9 RNPs are detectable at high levels shortly after transfection, and...plasmid based methods. One of the most common techniques for delivery of RNPs is electroporation (A in...the cytoplasm. In addition to the use of this technique in cell culture, it has also been applied to genome...combined with cell-type specific reagents in a technique known as nucleofection, which forms pores in the..., allowing for entry of a DNA template. Other techniques, such as lipid-mediated transfection (B in the...using this method for in vivo applications. This technique utilizes Cas9 proteins harboring receptor ligands...

...Sample preparation for these techniques sometimes makes it more difficult to detect protein tags using antibodies...availability of reliable primary antibodies for their detection, particularly when antibodies to the POI are non-specific...multimeric tags are frequently insufficient for detection when the POI is weakly expressed. As these epitopes...have no intrinsic visual signal, they cannot be detected directly in live imaging experiments. It is therefore... are both intrinsically fluorescent and can be detected with highly epitope-specific, fluorescently-labeled...can be easily expressed in the brain and can be detected through their direct fluorescent signals or using...antibody binding sites and primary antibody-based detection, smFPs create much brighter signals for weakly...

... commercially available mycoplasma detection kits that use detector cell lines, ELISA, fluorometry, or...Incidence, sources, effects, detection, elimination, prevention.” Cytotechnology 39.2 (2002). 75-90. Pubmed... lines, or infected personnel and can persist undetected; unlike infections with larger microbes such ...or bacteria, mycoplasma can be extremely hard to detect with levels reaching 108 cells per ml before the...or PCR-based approaches to detect even low levels of contamination. For those labs that prefer a more ...more economical mode of detection, Uphoff et. al. outline a do it yourself PCR-based approach complete with...alters microarray gene expression profiles.” Biotechniques 35.4 (2003). 812-4. Pubmed PMID: 14579747. 3...

...consists of prominent members of the bioscience and tech transfer communities who help guide and promote ...that had Platforms with a capital ‘P’ focused on technology ... has been critical to the Broad’s success ...biological questions, you need a tremendous amount of technology development alongside it.” Doench starting working...Perturbation Platform and encompasses a variety of technologies to functionally investigate the human genome...genome. Doench says he enjoys working on technology development under the Platform model at the Broad Institute...Institute because they are “not just developing technology for its own sake,” but work in true collaboration...history of RNAi left him a bit skeptical of the technology at the start of the CRISPR craze, but he says...

...microscopy technique for your study see Addgene’s blog post Which Fluorescence Microscopy Techniques is Best...fluorochrome is removed from all other detectors except its dedicated detector. To ensure proper compensation ...cell health, analyzing cell cycle stages, and detecting specific populations of cells in a complex tissue...sample (1). When combined with cell sorters, this technology can be used to isolate a specific subset of cells...fluorescence intensity.   What flow cytometers detect Cell size and complexity, measured using visible...directions. The degree of scatter is captured by detectors and provides useful information about the cell...emits a fluorescent signal that is captured by detectors. The strength of the emitted photons varies from...

...CRISPR 101" post. Next-generation sequencing technologies are the gold standard but they remain cost-prohibitive... assays (e.g. Surveyor® or T7E1) for mutation detection. While these methods paved the way for mutation...introduce digital PCR as an emerging validation technology. Digital PCR has several advantages over mismatch... require standard curves, and can theoretically detect a single target molecule in a sample. Digital ... real time PCR. These assays can be designed to detect either a donor sequence integrated through homology-directed... remainder of this post will focus on assays to detect NHEJ. These assays include a forward and reverse...between 200 and 500 ng of purified PCR product. Low detection limits: Digital PCR assays can accurately quantify...

...popular technique in neuroscience and developmental biology. This type of cell labeling technique involves...group developed the original iteration of this technology in 2007. Used to label neurons in the cerebellum...cerebellum of mice, the team appropriately named their technology Brainbow. Brainbow 1.0 used the fluorophores ...construct but do not undergo recombination can be detected by immunostaining of the truncated YFP. In Thy1...CMV-Brainbow construct at the embryonic stage. This technique could be used to target Brainbow to neurons by... promoter. Zebrabow is an extremely versatile technology as recombination can be induced in developing...cells. Tetbow optimizes Brainbow for current technologies Tetbow (Sakaguchi et al., 2018) overcame the...

...Einstein College of Medicine has recently published techniques to follow translation in living single cells....cells. However, before CRISPR-Cas9 these techniques were quite tedious and, even here, there is the question... symposium seeking inspiration to resolve the technical hurtle of monitoring protein turnover of specific...interesting talk by Prof. Erin M Schuman, a former Caltech researcher, now Director at the Max Planck Institute...Frankfurt am Main, Germany. She presented a novel technology, back then still unpublished, that allowed the...synthesized proteins with FUNCAT Professor Schuman’s technique applies metabolic labeling using a noncanonical...noncanonical amino acid, azidohomoalanine (AHA). In the technique, AHA is incorporated into newly synthesized proteins...

...Scientist at the Broad Institute, John Doench. CRISPR technology has made it easier than ever both to engineer...is true when designing gRNAs for using CRISPR technology – the “best” gRNA depends an awful lot on what...Gene knockout by NHEJ Gene knockout with CRISPR technology is usually accomplished by Cas9-mediated dsDNA... Created with BioRender.com. Two newer technologies offer an alternative to HDR for introducing ... another target C or A in the window. Another technology, prime editing (reviewed in Anzalone et al., ... although these are still early days for this technology, and the user may need to optimize numerous parameters...CRISPRa (activation) and CRISPRi (inhibition) technologies – a nuclease-dead Cas9 (dCas9) is directed near...

... permeabilization techniques — and so it may be necessary to try multiple techniques. For example, we... ability to detect endogenous levels of the target protein. It may only be able to detect the target protein... (or refute) that the antibody is selectively detecting the target-of-interest in your assay and sample-of-interest...must first confirm that a candidate antibody can detect the antigen of interest in the given protocol. ...supportive, the data must show that the antigen being detected in the intended application — here, flow cytometry...conclusively prove the ability of an antibody to detect the antigen of interest when expressed at endogenous... approach, you’re looking for an antibody that detects the target in the parental cell line, but not in...

... and can be used in a multitude of scientific techniques such as molecular cloning or molecular diagnostics...used for an array of different molecular biology techniques and experiments. End point PCR’s goal is to amplify... cloning to create a plasmid of interest, for detecting the presence of an insert or piece of DNA in a...reaction (qPCR) also known as real time PCR is a PCR technique used for measuring a starting DNA concentration...during the PCR reaction cycles is continuously detected via a fluorescent signal. The signal increases...concentration. The cycle in which the unknown DNA is detected compared to the standard curve can be used to ...right. ddPCR uses a water-oil emulsion droplet technology that fractions the PCR reaction sample into approximately...

...Importantly, conventional flow cytometry uses one detector to detect one fluorophore and therefore one cellular...fluorophores can “spill over” into detectors that are not the primary detector for that fluorophore. For example...single (primary) detector to identify a fluorophore, spectral flow cytometers use all detectors for all fluorophores...panels: five laser spectral flow machines have 64 detectors, allowing for the use of up to 64 markers in a...emitted by a fluorophore is guided to a single detector using a series of mirrors and filters. Importantly... FITC signal is spilling over into the primary detector for PE. In order to correct for this, compensation...the fluorescent molecules can be quite broad, detectors will usually only pick up a small portion of that...