...the journey of transformation and storage begins. Some samples are submitted as bacterial colonies in petri...inspected to verify that the samples arrived intact. Some deposits are sent from labs nearby in Boston and...packages have spent days to weeks in transit and some have been transported across oceans and continents...barcoded tube (see next section) and every time something is removed from this tube, the barcode on the ... received is immediately transformed and stored. Some samples need to be transformed with a micropulser...electric charge to help the DNA enter the cells. Some samples require a reduced antibiotic concentration...

...celebration, we thought it only fitting to share some stories of ways we’ve turned the available into ... coming from researchers outside the USA, having some materials available to order, yet not consistently... Sinapse Biotecnologia 2022 Brazil   In some cases, the distributors broadly helped increase ...means that our shipping team is always engaged in some form of problem-solving. When universal solutions...and its parts that helps the viewer visualize something that cannot be seen with the naked eye. In 2023...that we could identify the need for distributors in some countries. ‌We’ve developed systems to allow for...

...application, but it can certainly help you stand out.  In some fields, a cover letter is vital, particularly in...re applying to the specific company or lab. Take some time to research the lab or company via their papers...the beginning is often the hardest part, here are some examples to get you started! Dear Dr./Professor...skills for the position]. I am enthusiastic about [something that you love and is important for the job]. ... okay to start from a standard version, but take some time to make it fit the job you’re applying for....review grammar and spelling several times and ask someone else read your cover letter (as well as your resume...

...concern in healthcare today, but it doesn’t have to be some looming, abstract, convoluted scientific concept...not naturally resistant but has become resistant somehow overtime. One fantastic example of intrinsic resistance... drives microbes to develop survival mechanisms. Some microbes even make their own antibiotic compounds...such a rich diversity of microbes in the world if some didn’t develop, either through random mutation or...Drug resistance is not a black or white issue. Sometimes bacteria are resistant to lower concentrations... will be killed by a higher concentration, or sometimes more exposure time is needed for the drug to work...

...easy-to-understand set of PowerPoint slides. What are some of the more challenging parts of the job? With fast...goal posts” as a part of a case, consulting can sometimes be challenging. The fast paced nature of projects...of great output in a short amount of time can sometimes become cauldrons on the verge of boiling. On other...sciences, in business, in healthcare economics, or in some combination of these fields. I got my PhD a couple...performing in the market. Hence, as a PhD, an MD, or someone trained in any aspect of research in biology or...successful career in several different fields. While some people continue to enjoy the challenges of healthcare...

...acids. This is termed codon usage bias. In fact, some species are known to avoid certain codons altogether.... tRNAs bind to and deliver amino acids to the ribosome where they are incorporated into the growing polypeptide... Figure 1: An overview of peptide synthesis. A ribosome interacts with mRNA and charged tRNAs to build...critical for determining the rate of translation. The ribosome may slow and pause during elongation which may...synonymous mutations which have even been linked to some diseases. It has been reported that 5-10% of human...suggest codon optimization should not be used in some in vivo experiments and in biotechnology and therapeutics...

...the next steps on some common applications, join us as we get into the weeds on some of our favorite topics...always inspired by the new seasons, so we've made some big summer plans this year! New Blog Series We ... and tricks for those new to the lab, along with some behind-the-scenes peaks of Addgene’s people, culture..., video, and social media endeavors - in between some much-deserved vacations, sunshine, and beach trips...

...cold enough to keep some common types of reagents stable for years. In fact, some scientists speculate...sustainable practices in the lab, where I’ll list some easy but effective habits you can incorporate to...motivated and to feel accomplished. In addition to doing some of the quick things above, try joining or starting...your university could join. Find alternatives for some of the harmful chemicals in your lab. These chemicals...

...how they’re used, see some new techniques that take advantage of them, and give some tips on deciding which... organisms and their characteristic radiance for some time – there are records describing them dating ...realized that, although they are not bioluminescent, some types of corals and anemones are vibrantly colored...able to report new fluorophores like DsRed from Discosoma sp. and Katushka/mKate from Entacmaea quadricolor...

...to look for them? Or maybe you just want to make some really beautiful images? Multiplex IF may be just...similar to standard immunofluorescence, but with some extra considerations (Im et al., 2019; McLaughlin... antibodies from Addgene), or be prepared to run some extra controls and validate the antibody yourself...; this helps all viewers, especially if you have some complex patterns with several overlapping targets...experimental design. Like selecting an antibody, some of the requirements are similar to standard IF. ...being damaged by all the handling steps, or there’s some unexpected cross-reactivity. Keep in mind that you...

...nucleases are promiscuous. In other words, they can sometimes localize to unintended sites with sequence similarity...In this instance, a 5% off-target frequency is something to keep in mind, but likely will not have a significant...will likely be useful. Optimal gRNA selection With some CRISPR experiments, there is hardly any flexibility...bound by Cas are sequenced, has been tested with some success. The major con is that binding by Cas does...suitable. How to control for off-target events Sometimes there just isn’t a way around off-targets. Your...even when you’ve tried everything listed here. For some applications, this is ok, but it still must be controlled...

...irreproducibility with some solutions based on what I’ve learned. I hope you can apply some of these solutions... “reproducible” does not necessarily mean that someone has replicated it precisely. You can find a great... body works, generating new therapeutics, and sometimes formulating new biological concepts. However, ...ridiculous to expect that one should be able to apply someone else's new and incredibly useful technique simply...experiments. Biology is messy enough that by chance, some small number of the scientists will get a result...

... in the lab. The first post focused on streaming some lab waste for recycling- a process that’s integrated...well-integrated into our lab lives. Read on to learn about some creative approaches to lab sustainability, and find...single-use is now a popular theme in life sciences, some of these plastics are high grade and can be washed...markets. Rethinking our approach to lab waste is something we all need to do. Remember, it was only 50 years...practices to meet higher environmental standards. Find some easy action items on our other article in this series...

...about “group” or “peer” mentoring? How do I ask someone to be my Mentor? I have a Mentor – what do I do... led to the topic of “What makes a good Mentor?” Some of the Tweeted comments: “A good mentor has your...mind, rather than (or as well as) their own.” “Someone to act as your ‘cheerleader’ – inspire you.” “Also...think you can finish that book in 4 weeks!)” “Sometimes a mentor is a sounding board--a good mentor asks...want to find additional sources of wisdom. While somewhat lighthearted, the article does highlight key characteristics...

...also mutagenized proteasome subunit PSMB5 to find mechanisms of resistance to a proteasome inhibitor; CRISPR-X...bidirectional promoter, integrate at 11 specific chromosomal locations, and stably express the integrated ...however, these methods are not without limitations. Some, such as the well-known yeast two-hybrid assay are...protein interactions, whereas others may be too cumbersome to use in a high throughput manner or make it...for using the indicated antibiotics. In addition, some versions of the mAID vectors come with mCherry2 ... Yeast Vectors blog post concedes that there are some drawbacks with this technique. Great care needs ...are derived from the Ralser lab's own pHLUM minichromosomal vector, and besides being used for restoring...

... Project for some example proposals and consider applying for their accelerator for some hands-on guidance...about themselves--so don’t be shy and reach out! Someone from your alumni community is a great place to ... academia from graduate school or a postdoc. But some are also open to individuals from mid- to late-career...office yourself! Plus, at the federal or state level some professional societies host “Hill Days” where they...outreach groups. Grab a virtual coffee to chat with someone working in a science policy space you’re interested...

...geneticists are a lazy bunch who love to rely on the awesome power of genetic selection. Knock-in events tend...requires additional experimental manipulation. In some cases, injection of Cre is required, however the... design, delivery, screening, and more. Here are some highlights from the past few years:  Repair template...facing outwards, ensuring the PAMs remain on the chromosome after DSB formation, was most efficient (Farboud...of experiments imaginable in C. elegans. Equally, some of the “elegans” methods outlined in this post –...Wieland I, Gaul U, Forstemann K (2014) Efficient chromosomal gene modification with CRISPR/cas9 and PCR-based...

...Surveyor nuclease, save some of the duplex and run it as an undigested control. Sometimes the PCR products can...add 3 microliters of 5 mM star anise?” This is something I was expecting when I set out to make my first...lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells were then used to generate a monoclonal cell...editing across all the cells in the population, so some of the cells will still appear green despite having...Bunz F. Genetic knockouts and knockins in human somatic cells. Nature Protocols 2007; 2(11): 2734-46. PubMed...

...absence of a rigid cell wall. Here, I will cover some of the best ways to tackle mycoplasma contamination...stains like DAPI and Hoechst 33258 to stain all DNA. Some amount of expertise is required in this process ...become a habit before you even realize it. Here are some things to do: Wear a clean lab coat and mask while...culture is irreplaceable or very expensive, there are some ways to rescue the cultures. Elimination is a very...contamination and are the last alternative. There are some drawbacks while considering this treatment as the...permanent damage to the cell culture it is irreparable. Some cells may die due to longer incubation periods and...

...DNA found in bacterial cells, and someone new to plasmids may need some extra guidance to understand the...that replicate independently from the host's chromosomal DNA. They are mainly found in bacteria, but also...utilize plasmids across a wide range of applications. Some common plasmid types include cloning plasmids, expression...viral plasmids, and genome engineering plasmids. Some of the many things that plasmids can be used to ...