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...team of twenty excited undergrads. Team IISER-B’s idea for the iGEM 2019 is to take bacteria out of their...objective is to induce tolerance towards cold, we decided to focus on a special class of enzymes expressed...usually defined between a fairly nippy 20°C to a decidedly uncomfortable 45°C). Their more adventurous cousins...fruit juice clarification, meat tendering, and oligosaccharide synthesis. A major incentive to use cold-active...industry as it can potentially help minimize the incidental losses like spoilage and improve the productive...inefficiently that no industrial operation would ever consider them worth the trouble.  Enter ELSA: E. coli Learning...Acclimatisation To resolve this impasse, our team decided to create a hybrid system where we could incorporate...

...” has me considering the microbiome more often than not). If you’re someone who’s considering getting ...definitely enjoyed the lab!) and would therefore consider myself a neuroscience novice (newb). In this post...entire story-telling session organized by Story Collider and which featured a talk about the different ...Developing better AI programs gives neurobiologists new ideas about how the brain functions and vice versa. Even...getting into neuroscience from the mol bio side you’re sure to have many opportunities to learn about and develop...large data sets. Whether these data sets include videos of behavior, many many images of brain regions,...particularly interested in using AAVs because they provide a highly versatile means of delivering genes to...

...maintenance within an E. coli cell, but vectors can be widely utilized across many different cell types and each...budding. Yeast Centromere plasmids (YCp): These are considered low copy vectors and incorporate part of an ARS...are most similar to bacterial plasmids and are considered “high copy”. A fragment from the 2 micron circle...most commonly used selection markers in yeast and provides the element needed to overcome the auxotrophy ...as additional uses for said element. This link provides a more extensive list of yeast auxotrophic markers...auxotrophic selection in E. coli? Additional considerations HIS3 L-hisitidine no S. cerevisiae  no yes...experiments. MET15 L-methionine and overproduces hydrosulfide ions yes - Grow with methylmercury. S. cerevisiae...

... approaches a host cell surface are complex polysaccharide structures, many of which are easily varied...proteins could function as surrogate antibodies for identifying and characterizing sugars and sugar-containing... anything else, making it difficult to quickly identify possible gene function. In this post, I will describe...assay, including examples of RBPs successfully identified using this method. Step 1: Prepare a random fragment...transformants for appropriately sized inserts. We consider a library worth using if at least 40% of colonies...overnight), flip the membrane over and place it colony-side up onto a fresh LB plate containing an appropriate...onto a lysis solution-soaked filter paper (colony-side up) and leave for 1 h at room temperature. After...

...want to reconsider your approach, and try a different cloning strategy. Once you've identified potential...plasmid and all the steps you will have to take. Decide which technique you are going to adopt (i.e. restriction...digestion on an agarose gel. If the digestion does not provide the fragments of the size you expect, check the...region of the primer to 40-60 bp. It is always a good idea to perform primer optimization, especially if you...that, they can be open source (i.e. primer3) or provided by the major companies selling PCR reagents. Nowadays...Now that you've read our plasmid troubleshooting guide, you hopefully have some new strategies to tackle... the lab he develops new assays and processes. Outside the lab, he enjoy the outdoor in all the possible...

...-art methods of sex and gender analysis and 2) provides case studies with concrete examples of how gender...capacity of stem cells. Yet, very few researchers consider the sex of their cells when doing experiments—...between male and female cells. The result of not considering the sex of the stem cells was that their male... in Norway, the team realized they should also consider the sex of the stem cells. In this case, they ... many researchers I’ve talked to who’ve never considered the sex of their cells. For one thing, cells ...hard to determine. To remedy this, Shah et al. provide a table indicating the sex of the most commonly... there are numerous methodological articles to consider—we have loaded those we are aware of in our checklist...

...Instead they are detected with tyramide signal amplification. Tyramide signal amplification occurs when...are secreted from cells due to their IgGκ signal peptide sequence, making them easy to harvest from cell...when HRP converts a fluorescently labeled tyramide molecule into a highly reactive radical, which is then...Detection Method Uses Pros Cons Antigens HRP (P) Tyramide signal amplification of HRP Staining cultured...Enzymatic reactions on tissues can be cumbersome Tyramide reagents can be expensive Can only stain with ...of a nanobody that would make a useful RANbody, consider leaving a link to its sequence in the comments...immunodetection If you enjoy DIY lab projects, consider making you own DNA ladders with these plasmids...

... 1 we provide an example of such a comparison between cyan fluorescent proteins using widefield imaging... which usually are undesired properties. In the ideal situation, a fluorescent proteis should emit a stable...since different fluorescence imaging strategies use widely varying amounts of excitation power (Shaner et ... intense light (for a brief time), whereas in widefield imaging fluorophores are excited with relatively...measurements and hope this information can help guide your understanding of reported data. Photobleaching...Measuring photostability at realistic powers will provide better insight into the photostability in the intended...situation. Measuring photostability in living cells provides a much more realistic view of photostability. ...

...missense variants identified from 58 individuals with GNB1 disorder since the identification of this disease...and Accelerate Research Discoveries) Challenge provides grants for rare disease research. This year is...other symptoms as discussed in this educational video. “SLC6A8 belongs to a large family of solute channels...perform a functional screen in mammalian cells to identify any solute channels that can compensate for the.... “This proof of concept study will help in identification of new targets that could greatly enhance our...keratinocytes,” says Mellett. This would help the lab identify therapeutic targets in the future. Congratulations...will express these candidate genes in TS iPSCs to identify whether any rescue cell migration and contraction...

...secondary considerations to validation, application specificity, and experimental considerations - no other...you know which ones to try?  Read on and we will provide you with some advice on the antibody selection ...antibody first look at the application-specific data provided on the manufacturer’s website. Because of how ...has been validated in your specific application. Consider also that there may be variations within an application...section then a directly-conjugated antibody will provide better resolution as these immune complexes are...abundance on a Western blot, a secondary antibody is ideal as it will help to amplify the signal. Some applications...the species that the antibody cross-reacts with. Ideally you will find an antibody that has been validated...

...enzyme-bound hydropersulfide that is then transferred to thiol-containing acceptors. These hydropersulfides modulate...live cells. His work focuses on the enzyme hydropersulfide-propagating enzymes 3-mercaptopyruvate sulfurtransferase... susceptible to redox modification, the MPST hydrosulfide can be transferred to the roGFP2 cysteine thiols... to better understand the processes that happen inside the cell,” says Pedre Pérez. Figure 2: ...she studies opioid use disorder. Her goal is to identify cell populations in the brain that contribute ...neurons at specific firing frequencies. She hopes to identify how in vivo firing frequency of PV neurons affect...genetically encoded voltage indicators. Tian developed a video-based high-throughput screen for directed evolution...

...lentiviruses in parallel, this approach should be considered. Similar to the p24 assay, titering via measurement...When using qPCR based methods for viral titration consider using a reverse transcriptase mutant in a separate...carryover. Functional titer Functional titer is considered a more accurate method to quantify virus as it...procedure can take several months but should be considered in labs that will routinely be producing virus...virus or require precise titers. Additional considerations for titering lentivirus Volume and transduction...batch of virus for long term use, you may want to consider using an aliquot that has been subjected to a ... labs that routinely produce virus may want to consider the more accurate proviral integration assay. ...

... Time project provides a framework for these undergraduate research questions. The idea is straightforward...were generating huge amounts of data that could provide the basis for student projects in introductory .... Genome projects use computational methods to identify genes by their similarities to genes in other ...studies generally leave questions about gene function wide open. What if two seemingly similar proteins have...phenotype is corrected, students can be relatively confident that the gene from the test species has a similar...Education: A Call to Action, Washington, DC. 2. President's Council of Advisors on Science and Technology...about Synthetic Biology with BioBuilder Use Addgene Videos to Learn about Basic Lab Techniques High School...

...trials involve identifying a therapeutic gene, finding a means to deliver it and identifying a suitable route...technologies have allowed scientists to quickly identify underlying genetic causes of some human conditions...example, it was possible to transduce the T-cells outside the body and thereby correct the mutations using...productive infection. AAV vectors are engineered to provide additional safety benefits: they lack all viral...immunity. AAV vectors are also capable of infecting a wide range of cell types including but not limited to...through direct evolution, capsid shuffling, and peptide displays. These capsids will be selected both for...to other gene delivery tools that are under consideration, recent advancements in various AAV technologies...

...example at an anniversary or for Girls’ Day, offer guided tours to show people where the lab magic is happening...share your knowledge at local schools. Or work as a guide in local museums, zoos or botanical gardens. You...Addgene are open to guest contributions. If you decide to start your own blog, check out these two excellent...record a podcast or put some effort into making a video? Use your unique talent to communicate science. ... Guardian, BBC Breakfast or the Londonist. BBC provides a peak into their day-to-day business with popular...Co-Ordinating Centre for Public Engagement (NCCPE) provides on-site courses in public engagement for institutions...media skills, writing about your research and residential communication. If science journalism is the path...

...genome-wide library deposited by Leopold Parts. This library delivers 2 randomly paired guides per construct...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ... can both process crRNA and carry out the crRNA-guided DNA cutting, unlike other well-characterized Cas...to delivery constraints and scientists seeking a wider range of targetable genomic sequences could benefit...researchers sift through thousands of cells to identify cells exhibiting properties of interest. However...of Yellow Fluorescent Protein (YFP). The team identified a new YFP variant, mGold. mGold was found to ...reduction in performance compared to larger genome-wide libraries. Find the library here New from the...

...proteins had to be re-engineered for each gene, making wide-scale gene activation seem next to impossible. The... systems only require changing the programmable guide RNA. Gene activation by dCas9, also referred to ...specific sequences, CRISPR/Cas systems rely on a guide RNA complementary to the sequence of interest. Upon...VP64, a strong transcriptional activation domain. Guided by dCas9, VP64 recruits transcriptional machinery...of VP64 per each dCas9, SunTag uses a repeating peptide array to fused with multiple copies of VP64. By...two-component system dependent on gRNA design (SAM) or peptide design (SunTag). This streamlines its delivery,..., Crawford GE, Reddy TE, Gersbach CA (2013) RNA-guided gene activation by CRISPR-Cas9–based transcription...

... your video call.  Expect technical difficulties. To try and circumvent this, test out the video conferencing...development, and company culture.  It’s also a good idea to prepare 1-2 unique questions for each interviewer... homework before the interview. If you are not provided with a list of who will be interviewing you, ask... your audience. Connect your prior work or a new idea to the company’s area of expertise. This has potential...difficulties. Have your presentation on a flash drive and consider bringing your own laptop as a backup, if the company...the interview, if possible. Otherwise, read the video conferencing instructions you received from HR carefully...after completing each section of your talk. Some video conferencing software allows participants to “raise...

...just isn’t always ideal. But the increased flexibility helps even when it’s not an ideal site: moving several... that you hope is present next to the "perfect" guide sequence for your genome engineering experiment....editing landscape. The need for PAM-less editing   Guides for CRISPR are typically selected by the ‘quality...sequence, that of SpCas9, is NGG. Assuming all nucleotides are represented equally, that’s only a 1/16 chance...flexibility in gRNA selection and subsequent editing. Identifying PAM-free and -flexible Cas enzymes   The solution...more pan-genome PAM flexibility, you’ll want to consider one of several Cas9s that recognize NG PAMs. The...sought to address the PAM problem through structure-guided engineering of the PAM-interacting domain of Cas9...

... the last year for diagnosing COVID-19, aside from the widely used qPCR and antigen tests.  Labs including...Konermann S, Hsu PD (2021) Genome-wide, bidirectional CRISPR screens identify mucins as critical host factors...spike protein was developed based on 100 structure-guided spike designs and has improved protein yields and...During the 2003 SARS outbreak, Hyeryun Choe’s lab identified that ACE2 is the functional receptor for SARS-CoV..., and more. Recently, scientists used the genome-wide CRISPR libraries, Brunello (knock out) and Calabrese...expressing SARS-CoV-2 open reading frames and identified interactions of these proteins with human proteins...Months later, we partnered with Ginkgo Bioworks to provide synthesized SARS-CoV-2 expression plasmids available...