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...promoter Mature siRNAs typically range from 19–22 nucleotides and are 100% complementary to their target sequence...plasmid and call it a day. You need to include both sides of the complementary RNA duplex and the hairpin ...169)","rgba":"rgba(0, 82, 169, 0.75)"},"override_default_style":true,"text_hover_color":{"color":"#ffffff...time you need to knock down a gene, maybe you’ll consider using RNAi! References and Resources References... R. (2021). Reexamining assumptions about miRNA-guided gene silencing. Nucleic Acids Research, 50(2), ...

...so we’ve been busy updating our CRISPR guide. Our CRISPR guide is a comprehensive resource that covers...week at #AddgeneCRISPRweek. We updated the CRISPR guide! If you’re not into Twitter, we have some exciting...for planning your CRISPR experiment. For a quick guide to CRISPR, download our CRISPR Cheat Sheet....

... Summers, 1998). Multimeric plasmids are also considered to be less stable and more prone to being lost...other methods below should help in multimer identification.   Fig. 2: Gel image of Plasmid 61564...used to detect plasmid multimers. Some companies provide NGS whole plasmid sequencing services using long-read...within a plasmid and, therefore, allow for the identification of plasmid multimers. Capillary Gel Electrophoresis...laser-induced fluorescence (CGE-LIF) for detection and can provide data on the quantity of various topologies and ...

...insights into the process of getting an MD PhD, the inside scoop on developing new CRISPR tools, and the potential... : Alex introduces himself and explains why he decided to do an MD PhD - it's all in the family. 3:56 ...Biologically Inspired Engineering and why Alex decided to work there. 13:22 - 22:07 : Alex's projects ... Resources on Addgene.org Check Out Our CRISPR Guide Pages Find Ready-to-Use CRISPR Lentivirus Learn ...

...this concept in his microbiology lab course, he decided to add a bit of fun. He mixed together 10 E. coli...host strains affect transformation efficiency.  Aside from the Streak Plate Challenge, Westenberg’s course...plasmids here, and check out our streak plate protocol video on Youtube:     Additional resources on the...plasmids at Addgene Check out additional written and video protocols ...

... In vitro CRISPR screens in immune cells have identified many putative regulators of immunity. However...compatible with the introduction of unique molecular identifiers, which can improve the statistical power of in... of the pRDA_526 vector. Moreover, the screen identified putative novel regulators of CD8+ T cell responses...Context-Dependent Germline Knockout in Immune Cells Genome-Wide Screening Using CRISPR Cancer and the Immune System...

...some good guidelines to follow in order to ensure that people can quickly and easily identify what your...the insert. pBACKBONE-Flag-hGene-EGFP Other considerations  If your insert contains a mutation or modification...generally listed as the amino acid change and not a nucleotide change. The proper way to denote an amino acid...

...journals has risen steadily from 1995 to 2013. Considering citations to all articles, Anurag Acharya and...citations for older publications. (As an interesting aside, Nature recently reported that the most cited work...their own libraries and offering another way to consider and quantify the impact of scientific publications...find ways to capture scholarly work that exists outside of traditional scientific journals. Of course, ...

...puzzle fell into place, as they, taken alongside the insights provided by the studies in Tsien lab, covered...victoria (Figure 1) in molecular detail. On the underside of the “umbrella” of A. victoria there is a ring...1980s, another researcher, Douglas Prasher, got the idea that this new green glowing protein could be used...

...communication? It is important to identify your audience and consider their background. Social media is...contribution can either be an article, a short post, a video, or a piece of sci-art.  What are your top tips ...communication posts Sign up to receive our science career guide Learn about different careers in science communication...

...genome editing in human cells. The Feng Zhang lab identified a Cas12b from Bacillus hisashii, BhCas12b that...Microbiology Find the Mobile-CRISPRi plasmids Identification of Cas12a inhibitors Using a bioinformatics...a screening approach, the Jennifer Doudna lab identified three different Cas12a inhibitors. Some of these...sheet Resources on Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited...

...absence of certain markers on the surface or the inside of your cells. While proteins themselves can emit...fixable dye stains that react with free amines on and inside the cells and are thus typically used for distinguishing...vector.) In our flow experiment, we'll want to identify CD45+ cells (CD45 expression) and assess our transduction...should exclude APC-Cy7 from the rest of our panel. Besides some background cell death constantly happening...Compensation Resources on Addgene.org Addgene's Antibody Guide Ready-to-use antibodies in the Addgene repository...

...antibody, which recognizes the primary antibody and provides the method of detection. You’ll also want to use...is used to detect proteins on the surface of or inside whole cells. Antibodies - typically, primary antibodies...staining. This panel of antibodies is used to both identify the cell subsets of interest from among the mixture...This could be cells or tissue slices mounted to a slide, or whole organs. Because the tissue architecture...IMMUNOGENS, VACCINES, AND IMMUNIZATION. In: Immunology Guidebook. Elsevier, pp 17–45. https://doi.org/10.1016/B978...

...Stacey from Eastern Tennessee State University and provides a brief rundown of Sean’s top five smartphone ... lab life a bit easier. Since the post doesn’t provide an exhaustive list of all the apps out there, we...reasons: 1) I think phage are super cool and have considered getting a tattoo of my favorite molecular biology...to screen for proteins that could be useful for a wide variety of reasons. 3) I would love to see more ...

...Cre - A useful tool for neuroscientists AAVs are widely used by neuroscientists because they efficiently...interest is expressed.  A cell-type specific Cre can provide you with that control in a modular way. For example...only be expressed in neurons. Indeed, this AAV provides you with a means to specifically activate expression... viral vector blog posts for many more tips and ideas on how to use these fantastic tools in your experiments...

.... Although my interests are very clear, I had no idea how to translate them into real world careers. I...this new virtual world, I was worried about how my ideas would be translated through Zoom calls and Google...frequent Zoom meetings helped me communicate my ideas. I was able to express my visions for what my Instagram...with Quintin Marcelino who is Addgene’s Science Video Production Specialist and Arthur Guerra who is a...

... but if it is not possible consider a phone or video call. If you do decide to use Skype, make sure to... from other scientists doing interesting work. Consider this to be like any other research project. You...

...traction. One way to consider where the open access (OA) movement might be going is to consider some of the places...Access (YODA) Project, “a collaborative endeavor guided by aspirations to promote open science, ensure ...MOOC), with an explosion of courses and content providers and participation from many well-respected and...

...them remain uncharacterized. This represents a considerable barrier to furthering research and developing... an assortment of genes. Using patient data to identify mutations of unknown importance in cancer Previously... Center patient databases, this research team identified 1,049 perturbations in several genes spanning... 33 cancer types. The types of mutations they identified included gene amplifications and fusions, missense...

...Maureen Hanson’s and Rima Menassa's labs made the accidental discovery that laser treatment can photoconvert... Drosophila and mammalian cells, and it may find wide use in biological research. Photoconvertible fluorescent...great scientific discoveries, this one was made by accident. After exciting GFP-labeled plant cells at 405...variants used by Sattarzadeh et al., as well as a wide variety of other fluorescent proteins for your imaging...