...to track or measure the process you’re interested in. Depending on the process you would like to monitor... sensors’) are biological tools that monitor a process or detect a given molecule. The sensor component...over time. You can also use biosensors to follow processes such as protein or DNA binding, enzyme or transcriptional...  Detection Method Notes Quantification of a process Plate-based assays (plate reader) Detect the average...of cells (i.e. in a plate well). Use to track a process (e.g. enzyme activity) over time or under various...

...gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences...  Figure 1: Overview of the Western blot process. Starting with a mixture of proteins, it is separated...stained with antibodies, and visualized.   This process allows you to detect a single, specific protein...conduct the electric current. The wet transfer process is very effective in transferring a broad range...common readout for subcellular fractionations, a procedure that isolates proteins from different cellular... antibody specificity and antigen structure. Proceedings of the National Academy of Sciences 76:3116–3120...

...replenishing kits, is to find new ways to make lab processes more efficient. Being in the Boston area, we get...workshops and can use what we learn to improve processes in the Addgene lab. In this post, I’ll tell you...sample into many small components. Without proper processing, these small DNA strands can form concatemers...libraries from previously fragmented DNA.The general process for library preparation using this kit is: Fragmentation...that regardless of how much DNA we started the process with, we got great amplification of our sample ... for us as a company to constantly update our processes and get new ideas from learning experiences like...

...content. Regardless of the process, everything goes through an editing process and final check before publication...invite you behind the scenes in our content writing process, and to even get involved yourself. You probably...science, and our entire mission is to enable that process. Our repository of plasmids, AAVs, antibodies, ...recommendations. A few discussions in the review process led to quick dives back into the literature to ...and hit publish. Making content is an iterative process. We are constantly reviewing the resources we currently...

...biological sources of DSBs, and which need to be processed. Processing occurs prior to ligation as incompatible.... After the initial cut, the next steps in the process involve repairing chromosomal DSBs. It is important...step. Accordingly, NHEJ has a vast toolbox of processing factors, including polymerases (Pol μ and Pol...Although we do not describe these steps here, the processing of DNA ends tends to be the point where mutations...Non-Homologous End-Joining Really an Inherently Error-Prone Process? PLoS Genet 10:e1004086 . https://doi.org/10.1371...

...guide to our online data entry process. Figure 1: Begin the deposit process by clicking “Online Submission... our deposit spreadsheet?  Our plasmid deposit process can be broken down into four simple steps: Send... contact you if any issues arise during the QC process. As the depositing scientist, you will have the...information in a deposit spreadsheet. The data entry process On the Addgene home page, if you scroll over “Deposit... plasmid deposit process...

...to begin replication is a carefully controlled process balancing positive and negative regulators. Initiation...stable DNA-protein complex that will continue the process. DNA replication and regulation in bacteria Before...strands must be methylated before initiation can proceed. Newly synthesized DNA is primarily methylated ...replication of pSC101, a low-copy number plasmid, proceeds from a single origin of replication similar to...forks at either end of the origin. The helicase proceedes to unwind the DNA from these replication forks...the essential replicon of the plasmid pSC101." Proceedings of the National Academy of Sciences 80.21 (1983...

...cloning processes will require a PCR reaction or two to generate templates for the cloning process. Time... products via overhangs left over from the PCR process (the precise mechanism isn’t well understood). ...backbone. Once combined, the cut and ligation processes happen in a matter of minutes. Cost: Funding ...commercially available cloning kits can often make the process much more streamlined, because they will come with...reaction in favor of the desired assembly to make the process close to 100% efficient. Gateway cloning also offers...

...quality, we undertake a rigorous quality control process. In 2016, we launched our viral service to continue...bacteria snuck into the prep during the production process? These questions are especially important when ...potential contaminants. The analysis is a 2-step process using Geneious software: First, the individual ...CHRM4 gene. When contamination is suspected, we proceed to de novo assembly of the unmapped reads, employing...validate our method. We then extracted the DNA and processed the mixed sample in our pipeline (see Figure 2...answer your questions about our quality control process! The images in this blog post were created using...

... be tracked easily at each step in the storage process. After the original sample is barcoded, the lab...stocks, prepped DNA ready for our quality control processes - all of these samples have the 2D barcodes mentioned...a rack of tubes is dropped at any point in the process, we know exactly which location each should go ...step in the receiving, transforming, and storage process, we ensure that the sample tubes that we use match...Addgene’s secret to success. It helps us quickly process samples accurately and in a way that can be easily...deposited samples along every step in the storage process - from deposited DNA to transformed cells and finally...

...the grant development and management process or the whole process, including grant readiness, funding ... that likelihood. We provide expertise on the processes needed to successfully implement, manage and close-out... funds available and the increasing regulatory process for grants has fostered the demand for experts ...position that required some involvement in the grants process and managing programs that were grant funded. I...and experience through the candidate application process, the candidate’s ability to write a grant application...

...powerful resources for investigating any biological process. High quality genome sequence A high-quality sequence...understanding the gene networks that regulate processes from development to metabolism to fertility. Several...mutations in genes affecting diverse biological processes—not just development—are available from strain...Compared to the opaque mouse embryo, where similar processes occur over days inside the mother, the zebrafish...and explain how signaling pathways control this process. Additional zebrafish applications and conclusions... providing unprecedented views into cognitive processes like decision making (Kim et al, 2017). “Brainbow...

...because a process is working, doesn’t mean it will always be that way. Things change and processes often ...intentionally engage in a creative problem-solving process?  You may want to start by carving out time to ...holds the dual purpose of giving people time to process the issue properly while also discouraging everyone...is thrown out because they haven’t had time to process on their own. You may also find your meetings run...This is why perfectionists tend to also be procrastinators.  If you fall into this category, get out of...

...actually share many biological properties and processes with our own cells. Like human cells, yeast DNA...proteins and tissues to better assess developmental processes. For example, the Stainier lab studied the proliferation...making it ideal to study basic molecular biology processes. C. elegans are also quite amenable to genetic...investigate an array of biological systems and processes including the nervous system, development, cell...β-cells identified by HOTcre mediated labeling." Proceedings of the National Academy of Sciences 106.35 (2009... in freely behaving Caenorhabditis elegans." Proceedings of the National Academy of Sciences 113.8 (2016...

...team as a DNA Sequencing Intern. The application process When I first saw the comprehensive job description...leaving the interview hopeful to continue on in the process and I was excited when I received an email to invite...what my position entailed. The official training process was lined out to span the first three months of...the plasmid function. The steps of the training process gave me enough time to become familiar with the...there. At this point, I have seen several lab processes and am currently working on projects for the Science...

... enter the culture medium during subculturing processes or from the air if the cell line remains exposed... infecting mycoplasma. The primers used in the process are either species/gene-specific or universal. .... Some amount of expertise is required in this process because result interpretation might be difficult...by training eligible staff for the cell culture process and hence eliminate one of the causes of contamination...filtration. There is no 100% assurance that this process can avoid contamination but it’s a very good practice...cultures. Elimination is a very time-consuming process and increases the risk of secondary contamination...

...   While the VGS process isn’t new to Addgene, we are excited to share the procedures and Python scripts... vector (AAV) samples and created a simplified process, termed viral genome sequencing (VGS), to confirm...We recently published a description of the VGS process and use cases in Human Gene Therapy. Existing methods...substrates for tagmentation-based library preparation procedures. This finding eliminates the double-stranding...

...    Figure 1: In the isotype conversion process the variable regions of a parental antibody (blue...isotype (purple, mouse). Depending on the cloning process, all constant regions, just the heavy chain constant...isotype to that of a different group (Fig. 1). This process, called isotope conversion, gives users the flexibility...addition to increasing multiplexing capabilities this process is frequently used to reduce immunogenicity and...therapeutic potential of antibody medicines. In this process, antibodies derived from non-human species like...

...into a plasmid of interest. You can use similar processes to add overhangs to your insert of interest for... The steps following primer design and the PCR process itself are very similar to those outlined in our... with a few quirks specific to the PCR cloning process - please check out that post if you need a more... suited for analyzing the gene’s function. The process is shown graphically in the following figure, in...restriction digestion. As such, the later steps in this process are the same as those discussed in our restriction...

...Allikalt et al., 2020) Impact of labeling process: The process of dye labeling, such as conjugation with...beneficial when studying dynamic processes like mitochondrial dynamics. This process, involving mitochondrial ...specimens has offered illumination on biochemical processes that are crucial in the world of biological research...result in cellular toxicity due to the permeation process. Advanced technologies in fluorescent dyes/ligands...