... thinking, ‘wait, those terms mean different things?!’, you aren’t alone! We’ve noticed a lot of scientists...in. The differences They say the devil is in the details - and that’s where the differences in these... RNAi resources and go fishing! References and resources References Wang, T., Wei, J.J., Sabatini, D....selections The common goals The confusion in differentiating between a screen and a selection isn’t surprising... you would individually treat cells with many different drugs and then perform an assay to look at an ... No one wants to spend time screening hits that aren’t “real” or novel for their purposes… or worse yet...experimental question. Now that you understand the difference between the two, it’s time to check out our CRISPR...

...findability” of the information on the site. Different users have different strategies for finding what they need...the overall user experience of our website, the different website navigation paths, the organization and...software team tailored Elasticsearch to support the different usage patterns the larger team identified. The...our catalog of materials and services is quite different from the way it searches through our educational...information in our search indices in order to support different kinds of matches: exact words, partial words, ...or numerical identifiers. We also combine the different terms in a query in a way that allows users to...Weighting We determined the relative importance of different bits of information for a given resource, and ...

...overexpress or knockdown a gene, you might need different plasmids. Harvard’s DRSC/TRiP Functional Genomics...varying expression levels of your transgene in different fly lines. Also, if there is a chance you need...transgenes in one fly, having the transgenes in different loci allows for genetic recombination so that ...for direct comparisons between lines expressing different variants of YFG because the transgenes are found...remember that you’ll need to integrate them all at different loci. Fortunately, there are many docking sites...docking site is located). This amounts to a small difference of two or three weeks. 3. How would you like ...experimentation. Drosophila are amenable to many different kinds of experimental assays depending on your...

...between Cas9 enzymes from different species, explaining why they recognize different guide RNAs. Deep in the...Other CRISPR enzymes like Cas12 have different structures and different uses. In all cases, structural biology...includes most of the bridge helix. Created with BioRender.com.   Cas9 has two major lobes, called NUC ...guide RNA (PDB ID 4ZT0), shown opaque (left) and transparent (right). The RNA guide sequence is extended as...target DNA (PDB ID 7S4X), shown opaque (left) and transparent (right). Some nucleotides of the non-target DNA...from the same angle to highlight conformational differences. HNH, red; RuvC, orange; CTD, yellow; REC lobe...; DNA, gray; PAM site, yellow. Created with BioRender.com.   The exact conformational changes that...

...containing many different promoters, RBS’, coding sequences (CDS), and terminators of varying strengths. These ... developed for the study of enhancer function, strength, and binding (e.g. DHS-seq, FAIRE-seq, and CHIP-seq...variable availability of enhancer sequences in different chromatin states. Alexander Stark and colleagues...transcription from the minimal promoter and its strength can be determined from its enrichment over input...process of cloning, combining, and testing these different parts can be quite arduous. The cloning process...that control many cellular processes such as differentiation, proliferation, and apoptosis. In humans, mutations...To facilitate innovative cancer research, the Reference Reagents Group of the NCI RAS Initiative, led ...

...ligation mix in different bacterial strains (stbl2, NEB stable, etc.), and/or use a different temperature ...want to assemble together in a combination of different ways according to your cloning strategy: Restriction...buffers and temperature are compatible between the different enzymes. Always check the fragments size of your... There are multiple ways you can assemble the different parts of a plasmid based on the cloning strategy...attention in the areas of ligation between the different fragments If you don’t get any positive clone...might want to reconsider your approach, and try a different cloning strategy. Once you've identified potential...Nature, and other journals.   Paolo Colombi is currently the product development scientist at Addgene. ...

... These each have different goals, formats, and processes, and they provide different amounts of access...research during his PhD studies.  References and Additional Resources References ASAPbio, Preprint Resource ...like publishing a research article, presenting a conference poster or talk, or submitting patent or grant..., and increase their impact. Open Access and Transparency Another major benefit of preprints is their ...Going to present some unpublished results at a conference? Post your working manuscript as a preprint too... work too openly too soon or simply a lack of awareness of the benefits of preprints. Some worry about...be prone to biases or errors, with a lack of transparency in most review processes. Preprints and their...

...want to load 1.0 μg in 10μL, make 1.1μg in 11μL. Reference Page Top Index...

...mini-prepping and verify insert by sequencing. Reference Page Top Index...

...section, teams are assigned different groups of three genes selected from different parts of the Met pathway...first select a multi-gene pathway to study and a reference organism where deletion strains are available....Students next transform deletion strains from the reference organisms with the overexpression plasmids. Finally...species has a similar function to the gene in the reference species. We have been using yeast models (3, 4...yeast, Saccharomyces. cerevisiae, makes a great reference organism for the project because it is non-pathogenic.... Over time, the two genomes have become very different due to a whole genome duplication and extensive...cloning to insert the coding sequences for ten different MET/Met genes (Figure 1) from the two yeast species...

...engage broadly with diverse communities, to raise awareness about where science is and where it is going, ...Personal Genetics Education Project (pgEd), where I currently work, strives to broaden the conversations about...Harvard Medical School, has a mission of raising awareness and starting conversations about both the benefits...and epigenetics. So, I sought out a number of different opportunities to broaden my training beyond the...major part of our everyday work is to liaise with different organizations and individuals, to identify ways...collaborating, and to organize events that bring different voices together to talk about genetics. In addition... such as scientific, educational or industry conferences, as well as schools, community events, churches...

...of the Canton Nucleic Acids Forum (CNAF) 2015 conference invited me to attend and speak about Addgene’...attended by over 300 Chinese scientists eager to hear current findings in nucleic acid science. I was delighted...Thoughts on Guangzhou and the Science City The conference was located in the modern, relatively new “Science...national science and technology park, it enjoys preferential governmental policies for a high-tech development... which I could identify. There were dozens of different mushroom varieties alone. At least I could tell...requires an interdisciplinary team.  Day two of the conference was kicked off by Nobel Laureate (and Addgene...circRNAs do yet or how they do it. Of course, No conference would be complete without CRISPR Michael Terns...

...to learn how to adapt to different situations and, in particular, to different styles of leadership. As...must accommodate many different types of people in order to harness their strengths and work effectively...to learn more. Having met many people at that conference, I later became involved with the NPA beyond ...within the committee, I became Vice-Chair when the current Vice-Chair stepped down. When the Chair then stepped... the Committee which I then led for 5 years.  Different leadership styles Being an effective leader requires...requires you to understand how to work with many different types of people both above and below you. During...relationships with these individuals who often had different styles from their predecessors. Changes were implemented...

... or looking to improve your current methods, understanding the differences can help you make an informed... Resources References Shaw, G., Morse, S., Ararat, M., & Graham, F. L. (2002). Preferential transformation...make the virus. Viruses can be produced in many different cell types, spanning both prokaryotic and eukaryotic...However, some viruses can be produced in multiple different cell types, including plants, mammals, and insects... transfer plasmid. The transfer plasmids have different packaging capacities depending on the virus. AAV... which system to use will come down to which differences are most important to you — virus type, cost,...this post has provided some insight on these differences in viral production and will help you decide ...

...first successful attempt to render fixed anatomical preparations transparent was done by Walter Spalteholtz... Pubmed. J. Sharpe, U. Ahlgren, P. Perry, Hill B, Ross A, J. Hecksher-Sorensen, R. Baldock,  D. Davidson...microscopy coupled with techniques that make tissues transparent are enabling microscopes to visualize the cellular...spinal tract, CST, spinal cord and cranial nerve afferents labelled with a AAV8 expressing eGFP under the...sample and captured the reflected light using a different objective lens oriented perpendicular to the plane...of clear specimens and for in vivo imaging of transparent organisms. Advances in tissue clearing Similar...Find plasmids from the Tsoulfas lab here.     References: H. Siedentopf, and R,. Zsigmondy.  (1902) Ann...

...presenting your research at conferences such as SACNAS and ABRCMS. These conferences have a lot of programming...her on Twitter @EleanorWang.     References and resources References Canning EA, LaCosse J, Kroeper KM...new therapies can be developed and applied. The current pandemic serves as a striking example of the applicability... is a supportive environment—it makes all the difference in a lab experience. “Mentorship” was a bit of...interested in doing research and attending research conferences. The Barry Goldwater scholarship is also a national...graduate programs, recruiters will attend these conferences to answer questions and offer application fee...explore! Addgene has an entire page dedicated to different career options from science policy to science ...

... This preferential tropism for different brain regions is largely due the ability of different serotypes...tissue? What is commonly known is that different serotypes have different tropisms but, as you can see, AAVs...there are marked differences among serotypes in their efficiency at transducing different brain regions....serotypes to infect different cell types. In the CNS, it has been shown that there is variable transduction...skeletal muscles, and the central nervous system. Currently, AAVs are being used to deliver CRISPR/Cas9 but...transduction efficiency, and safety profiles should strengthen its appeal for the scientific and medical communities...movement. Follow her on twitter @katrinaboards.   References 1. Russell, David W. "AAV vectors, insertional...

...across multiple teams or to stay abreast of current trends in the literature, sampling a breadth of work...One thing many scientists enjoy is discussion of current new and exciting literature. As I transitioned ... had concerns that I may not have time to stay current or enjoy that discussion. Luckily, one needn't ...science, researchers of all varieties must stay current with literature and emerging technologies to remain...meeting amongst scientists that aims to help stay current on new and emerging work by collaboratively discussing...the meeting settles in and people get the time recurrently carved out. This notice typically should include...the same figure by figure organization with a different person responsible for each figure. Others still...

...packaging; however, they are different viruses and thus use slightly different isoforms of these packaging...Generation 3rd Generation Viral Production FAQ References Glossary Lentiviral Components To increase the... contain an additional deletion in the 3'LTR, rendering the virus “self-inactivating” (SIN) after integration...packaging system. For a comparison of the key differences between the 2nd and 3rd generation packaging ...Questions (FAQ) about Lentiviral Plasmids What is the difference between 2nd generation and 3rd generation lentiviral... 3rd generation packaging system. What is the difference between a lentivirus and a retrovirus? Lentiviruses...retrovirus. From an experimental standpoint the main difference between lentiviruses and standard retroviruses...

...these tools can be to use. AAVs, which can target different cell types depending on their serotype, are particularly...93,491 viral vector preps with researchers in 45 different countries — and as of 2024, we’ve distributed ...the vectors in Addgene’s catalog are offered in different serotypes. In fact, the 879 viral vectors in our... CRISPR, where we dove fairly deeply into the different types of tools scientists were using/depositing... is less amenable to such analysis, due to its inherent bias: Addgene uses a prioritization process to...service. But why are their request patterns so different? It could be a matter of timing, as jGCaMP7 was...from requestors and depositors, and identifying trends in our own repository data, giving us significant...