...unmethylated sites in the chromosome. The more stable endonuclease can then cut the chromosome leading to cell ...Applications of ccdB in plasmid cloning Perhaps the most widely used application of CcdB is GatewayTM cloning...protein-based toxin-antitoxin systems. One of the most well known protein-based toxin-antitoxin system ...-Kulka, and Gad Glaser. "An Escherichia coli chromosomal" addiction module" regulated by guanosine [corrected...

...CRISPR systems were first discovered in bacteria, most CRISPR-based genome engineering has taken place ...finding suggests that the system may be functional in most Enterobacteriaceae. An update to this system, called... RNA interference does not function in bacteria, most efforts to regulate gene expression were limited...adenine base editing in Streptomyces Some of the most industrially and clinically relevant bacterial species..., yet genome engineering remains challenging for most organisms in the genus. Work from the Sun lab demonstrates...base substitutions respectively. Substitutions seem most efficient when using a nickase version of Cas9, ...

...have to be this way, if you know what you need. Most often, early career scientists aren’t trained to...it will take to publish based on these findings. Most often we suggest estimating the time it will take...strong negotiating tactic, and puts you ahead of most start-up P.I.’s. It doesn’t have to be complicated...condition and availability of the equipment is of utmost importance. Conducting a successful lab start up... and know how to decide which components are the most critical to your research, negotiations can be much...

...are the most common examples of the E. coli B strain. Common E. coli strains used in the lab Most of the...that make these features possible are present in most commercial strains, especially mutations that make...Streptomycin Hybrid of E. coli K12 and E. coli B (mostly K12, though), common lab strain for cloning and...covering E. coli and others are model organisms! Our Most Popular Plasmids 101 Posts: Origin of Replication...

..., 2016). Despite the almost universal functionality of the CRISPR system in most organisms, some plant-specific...include the ubiquitously expressing cauliflower mosaic virus 35S promoter (CaMV 35S) or ubiquitin promoters...2020) and plants viruses (barley yellow striate mosaic virus (Gao et al., 2019) or sonchus yellow net ... associated publications or protocols to get the most out of the plasmid you chose for your experiment...

... house mouse (Mus musculus) Let’s start with the most preferred mammalian model organism of many researchers...Morgan discovered that genes were found within chromosomes using the fruit fly far before we even knew that...Like human cells, yeast DNA is packaged into chromosomes and about 23% of yeast genes have a counterpart...that overexpress the toxic protein α-synuclein (the most common protein found in lewy bodies), have been ...al. "Efficient gene transfer in C. elegans: extrachromosomal maintenance and integration of transforming...

...fluorescent protein to study its function is one of the most popular applications of fluorescent proteins. These... encoded by DNA. Since researchers can generate almost any DNA sequence in the way that they like, the...vitro, ultracentrifugation or gel filtration is the most common analytical method. However, these may not...- and C- termini of your protein of interest The most straightforward fusion protein is made by connecting...The recent revolution in genome editing methods (most notably CRISPR/Cas mediated HDR) enables the tagging...

... approaches to record keeping. Top priorities in most cases include IP protection, publications and funding... in depth and provide the solution that fits the most labs, our team conducted a large study which focused...ease of use would influence their ELN choice, with almost 80% rating it as very important. One comment reflected...support your lab’s growth in the near future? As most payable software solutions have user-based pricing...company, site licenses might be available as well. In most cases this applies to universities. How are you ...

...recognize that there are different types of conflict. Most people perceive any conflict as inherently bad, ...Makes You a Good Conflict Resolver? The first and foremost sub-skill to have here is pretty intuitive: You...empathy does not mean you like everyone you meet. Most people can’t maintain that level of positivity past...Of the tools I’m mentioning here, this may be the most controversial, so I would like to qualify it. If... accommodating, if you find yourself gravitating most frequently towards these approaches, you may need...

...an essential vitamin for life Although biotin is most commonly known for its role in molecular biology... for life. Biotin is only synthesized by plants, most bacteria and some fungi, thus mammals must obtain... vitamin through external sources. Humans absorb most of the biotin they need in the small intestine from...specific post-translational protein modification. Most organisms have less than five enzymes in the entire...manipulated for a scientist’s intended purpose. Two of the most common uses of biotinylation in the lab are protein...

...in 2013 found that Gibson assembly was one of the most commonly used assembly methods (Kahl 2013). However...in which very large fragments of the bacterial chromosome (up to 100 kb) were specifically cut out via ...contain regions of homology with the bacterial chromosome fragments. CRISPR cutting was used to circumvent... the need to PCR amplify the fragments of the chromosome, which was technically challenging. Other homology...Ebenstein Y, Zhu TF. Cas9-Assisted Targeting of CHromosome segments CATCH enables one-step targeted cloning...

...the way. That’s what makes it so wonderful. The most challenging part of jumping into this career path...explainer of the who, what, where, when, why and most importantly, how, of a science policy career.  Who...engagement with your local or state governments. Most communities have opportunities to join commissions...do science policy? I would argue that this is the most important question that you can explore. The answer... important skills for the profession. One of the most important skill sets for a science policy career...

...research is rooted in the CNS, my focus will be mostly on the brain, but many of these tips apply to all... estate and using AAVs have in common? The three most important items when looking for a house and choosing...Well, an AAV user must determine which serotype is most efficient at transducing the cell type under study...influences on overall cell function. AAVs are the most appropriate vehicle for this type of research, as...zinc finger nucleases, and many others (12). The most attractive potential of AAVs is their ability to...

... pass on to you to make sure you are getting the most out of the data you get back from your sequencing...controls help you properly visualize your results.The most important of those is to always look closely at ...miscalled peak, don’t be shy. Feel free to edit it. Most chromatogram viewing programs (even the free ones...sequence.   I hope these tips will help you get the most out of your DNA sequencing results and to troubleshoot...

...become. One thing is for sure, as is obvious from our most requested plasmid, SpCas9 is still going strong ...editing experiments. So, without further ado, the most request plasmid in 2016 was... pSpCas9(BB)-2A-Puro... lines. Cas9, of course, is the key component of most CRISPR systems and generates double strange breaks...name are some additional features. The first and most imporant is that, upstream of the Cbh promoter, ...

...Centre for Regenerative Medicine (CRM), Edinburgh. Most commercial antibodies do not work in pull-down assays...diverse biochemical tasks in the cell. One of the most popular approaches to study protein interactions...methods (electroporation, lipofection etc) is the most crucial aspect of knock-in experiments. You’ll need...depends on many factors. Some of these include: Most importantly, the crRNA should cut near the insertion...with custom synthetic cr:tracrRNAs (csRNP) is the most efficient and convenient choice for epitope tagging...

... network of relationships early and often is the most effective tactic a scientist in training can adopt...professional (and personal) relationships. However, most job seekers will and should apply for jobs posted...get help if you are having trouble preparing your most excellent resume. 2) Follow job boards and check...letters that expound on bench skills and experience. Most Addgene PhD level job descriptions clearly describe...

...helix, fragmenting plasmid DNA, PCR product, or chromosome. One of the main causes of degradation during...endonuclease I encoded by the gene endA. Nowadays, the most common E. coli strains used for plasmid amplification...preparation stabilizing the plasmid, and still allowing most downstream processing. On the other hand, nuclease...where you plan to store it and for how long. The most common solution is to keep your plasmid at -20°C...

...alteration. Molecular cloning techniques are some of the most widely used techniques in the biological sciences...Although each type of cloning has its advantages, most scientists still encounter several struggles with... overhangs, insert and vector purification, and, most importantly, excessive time requirements. FastCloning...polymerases like phusion. An 18-cycle PCR leads to the most colonies after transformation. When a 1:1 vector...

...less waste.  Most communities seem to have recycling programs and it seems to me that most Addgenies make...advent of techniques like PCR and DNA synthesis. Most molecular biologists bought the pricey stuff in ...only Addgenie who has a thrifty streak.  One of the most accessed blogs on BitesizeBio is the protocol for...