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We narrowed to 996 results for: ran

Showing: 441 - 460 of 996 results

  1. 5 Uncommon Ways to Gain Career Skills During an Uncommon Time

    Type
    Blog Post
    ... and not in academia is run by grant funding. Learning to write grants quickly, efficiently and effectively...apply to or offer to help with lab grant writing. You may not get the grant or your PI may edit everything...thinking about future career options. 5. Write a grant - even if you don’t need one A lot of science in...valuable skill. There are many different kinds of grants that you can apply for. If you are in a lab, your...Here are some sites that have compiled lists of grants: Science: Where to Search for Funding The Ohio...For Your Research University of Florida: Training Grants and Other Funding Opportunities L’oreal USA For...

  2. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...method is transfection of short interfering RNA (siRNA). Knockdown caused by siRNA is also transient, and ...interest. This approach often involves the transient transfection of expression plasmids in a given cell ...commercially available. Various proteomic and transcriptomic datasets can be used to select a candidate ...antibody should be able to detect even in cells transfected with low efficacy. But since there can be large...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...workshops. Let’s use TIM-1 (Single pass type-1 membrane protein), which has been designated CD365 in the...the antibodies both recognized the target when transiently overexpressed in CHO cells and displayed similar...

  3. Cultivating Community Science at BosLab

    Type
    Blog Post
    ...2015, there were six of us. We were initially strangers but we all shared a passion for DIY Biology and...December 2015 we held our first intro to bacterial transformation course (read our blog post on it and to see...consisting of over twenty members and volunteers. Transforming citizens into scientists Anyone can become a...dedicated time at BosLab, he became our bacterial transformation guru and instructor for our intro to biohacking...biohacking courses. Our members pursue a diverse range of projects at BosLab. Some are entrepreneurs using...RNA with small molecules. Other members’ projects range from hacking probiotics to filming educational lab...BosLab. Once he optimizes plant cell culture and transgenesis he wants to share his knowledge with the community...

  4. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    ...covalently joined, scarless DNA – perfect for transforma­­­tion or as a PCR template. Some of the applications...by using NEBuilder which requires only a single-stranded synthetic oligo to generate a single-guide RNA...RNA (sgRNA). Using only one single-stranded oligo, instead of the more typical two oligos, is more cost...annealing step usually used to create a double-stranded oligo. Generating an sgRNA construct becomes as...target region. Assembled plasmids can be directly transformed into E.coli for plating, screening and generation...complexity, is achieved with synthetic oligos with a randomized region between the regions of homology. Producing..., generating expression cassettes for yeast transformation can take 5 days. NEBuilder shortens that time...

  5. What Good Citizenship Can Do for Reproducibility in Science

    Type
    Blog Post
    ...differently. Communication comes in many forms—from transparency about data to allow others to see what you actually...communication and sharing fits together to make for a transparent and collaborative research environment. Journals...Currently, most of the burden for pushing for more transparency in science communication falls on the shoulders...developed STAR Methods to improve the structure and transparency of methods sections, including clear information...researchers are not strongly rewarded for being transparent in their communication or making their data and... to a large extent, hold the keys to getting transparency and communication further up the collective ...management plans and for evidence of sharing for grant renewal—but there is clearly room to do more. If...

  6. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ...Cas enzymes) cleaves both strands of its dsDNA target in cis and has no trans activity. AsCas12a, on the...cleaves ssDNA in trans. In sum, CasPEDIA documents the target substrate (DNA/RNA and strandedness) along with...Cas enzyme: Nuclease properties (CasID): Cis/trans activity, PAM requirement, gRNA, and multiplex ability...features: CasID CasPEDIA will display both the cis and trans activity of each entry. For example, SpyCas9a (one...can manually enter details to refine your search (trans activity, multiplex ability, gRNA design, etc.).....32942/X2C31F     Makarova, K. S., Wolf, Y. I., Iranzo, J., et al. (2019). Evolutionary classification...

  7. OCaMP: A New Calcium Indicator for Neural Imaging

    Type
    Blog Post
    ...various teal, green, yellow, orange, and red fluorophores. The spectra for mOrange2 (scaffold of OCaMP) are...are shaded in orange. Data from FPbase.org. Scientists have developed calcium indicators spanning ...complement these tools, we recently developed a new orange calcium indicator, OCaMP (Aggarwal et al., 2025...2025), based on the fluorescent protein mOrange2 (Figure 2). Figure 2: OCaMP structure and mechanism...CC-BY-NC-ND 4.0 International license. Why orange? OCaMP’s emission peak at 565 nm fits neatly between...for two-photon imaging with newer lasers in the range of 1000–1080 nm. These industrial fiber lasers are...References Aggarwal, A, et al. (2025). A sensitive orange fluorescent calcium ion indicator for imaging neural...

  8. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    ...lactose analogue, IPTG (isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate, IPTG binds...x-gal (5-bromo-4-chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble blue pigment...α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a functional...site (MCS) into the α-peptide (represented as an orange wedge in the figure on the left) and inserted it...produces pigment from x-gal (included in the transformation plate medium), turning the bacterial colony...IPTG is also included in the medium to ensure transcription of the lac operon.   Tips for blue-white screening...screening Use a good control: Transform the backbone plasmid without insert. All colonies on this plate...

  9. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...students first transform the S. cerevisiae met deletion strains and then isolate transformed strains on media...organism into overexpression plasmids. Students next transform deletion strains from the reference organisms ...to a whole genome duplication and extensive rearrangements in the S. cerevisiae lineage (7). In our class...class research project, students transform S. cerevisiae met deletion strains with overexpression plasmids...expression of the plasmid-encoded MET gene allows the transformed strain to grow in the absence of methionine (... to select yeast that have been successfully transformed with the plasmids, because the S. cerevisiae ...of MET/Met genes from the plasmids allows the transformed strains to grow in the absence of methionine....

  10. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...split-vector systems’ by separating viral genes and transgene expression across several plasmids. The most commonly...separate envelope and packaging plasmids as well as transfer plasmids. This concept ensures safe handling and...Gamma-retroviral vectors have a relatively high transduction efficiency, but in contrast to lentiviral vectors... are very versatile tools as they are able to transduce dividing and non-dividing cells. More detailed...Gamma-retroviral vectors Popular gamma-Retroviral Transfer Vectors   Expressing your gene of interest pMKO...events and is widely used in the field of mouse transgenics. Experimental uses include: Cre-dependent gene... lab. Learn more about CRISPR plasmids for transcriptional inhibition. Retroviral Plasmid Kit Retroviral...

  11. Streaking for Single Colonies: The Streak Plate Challenge

    Type
    Blog Post
    ...and spread over the second quadrant. The process continues until all quadrants are used, hopefully revealing...students transform them into different strains of E. coli,” says Westenberg. During the transformation experiment...plate, first the bacteria are inoculated over a quadrant of an agar plate. Then, a stick is dragged through...revealing single, isolated colonies in the last quadrant after incubation. The challenge is usually successful...genetic backgrounds of the host strains affect transformation efficiency.  Aside from the Streak Plate Challenge...

  12. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ...non-selective anterograde transport to a lack of genetic control. ATLAS (anterograde transsynaptic label based on...of the color spectrum more powerful, the lab of Francois St-Pierre recently developed a pair of new yellow...in other plant species, thanks to the broad host range of TRV. Find TRV-TnpB plasmids here! Weiss, T.,...Viral delivery of an RNA-guided genome editor for transgene-free germline editing in Arabidopsis. Nature Plants...lung cells via airway administration. AAV11 can transduce neurons for retrograde labeling, making it a powerful...allows VAMP2-At to remain on the presynaptic cell membrane while AF, fused to a payload, is released into...). ATLAS: a rationally designed anterograde transsynaptic tracer. Nature Methods, 22(5), 1101–1111. https...

  13. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...growth rate/transfection efficiency is not known, this is easily achieved by splitting transfected cells into...specific cell line, and its transfection efficiency. We often trypsinize the transfected cells on the day after...after transfection into plates containing selection agent at different densities, then dispose of the plates...under-expressing clones. If GFP-encoding cDNA is co-transfected at a favorable ratio to the gene of interest ...such as the Viaflo from Integra Biosciences) to transition from a 48-well cell plate to a 96-well assay ...expression characteristics from a population of transfected clones. The technique is applicable to a wide...

  14. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...epithelial origin. Suppressing function, creating tolerance, and evading recognition are a few methods by ...regulatory cells to suppress function. Creating Tolerance T-regs in the tumor microenvironment produce CTL... This in turn regulates the expression of the tolerance-producing antigens enunciated above. Finally, ...released from the microenvironment help create tolerance beyond direct proximity to the tumor; chemokine-aided...relapse. Chemical and radiation therapies cause a transient shutdown of the immune system, hampering its ability...microenvironment. Antibodies that block cell-surface tolerance-producing proteins are one of the most commonly...cells to knockout proteins involved in immune tolerance. Recently, China spearheaded the first ever CRISPR-based...

  15. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...for a growing organism to make the complicated transition from a clump of cells to a fully differentiated...cooperate to cause a higher-energy electronic transition in a fluorescent molecule (see figure to the ...past 15 years efforts have been made to create transgenic mice expressing fluorescent protein reporter ...microscopy enabled scientists to see how chaotic and random the scanning was and how fast T cells could jump...Blue-green fluorescent proteins like EGFP Yellow-orange fluorescent proteins like TagRFP, tdTomato, DsRed...tdKatushka2 (Drobizhev et al. 2011) The commonly used orange and red fluorescent proteins are excited by 750...essential role for neutrophils in leishmaniasis transmitted by sand flies." Science 321.5891 (2008): 970-...

  16. Kwabena Duedu on Public Health, Biofuels, and Doing Science in Ghana

    Type
    Blog Post
    ...applicant to the Addgene, Seeding Labs Plasmid Grant. For this grant, we’re working with fellow nonprofit, Seeding...about the Addgene, Seeding Labs Plasmid Grant Read about Plasmid Grant Recipient, Gladys Kaba Resources on...their research. Kwabena came across the plasmid grant in Seeding Lab’s Newsletter and recently visited...Kwabena applied for the Addgene, Seeding Labs plasmid grant. 7:12 - 7:11 : Kwabena's work in Christopher E. ... recipient of the Addgene, Seeding Labs Plasmid grant and is working on a variety of projects in Ghana...

  17. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ... the P and V proteins of Simian virus 5 (SV5) (Randall et al., 1987). The epitope (GKPIPNPLLGLDST) has...10.57733/addgene.l0ulu4. Hanke, T., Szawlowski, P., & Randall, R. E. (1992). Construction of solid matrix-antibody-antigen....org/10.1099/0022-1317-73-3-653. PMID 1372038. Randall, R. E., Young, D. F., Goswami, K. K., & Russell...functional properties, and is amenable to a wide range of biophysical assays (Kroupova et al., 2024). They...using mCherry for selection of sgRNA plasmid transformants, they avoid M. abscessus’ high rate of spontaneous...workflow in M. abscessus. A) Sth1Cas9 mediates double strand break at the target site, which results in either...indels after DNA repair. B) M. abscessus is first transformed with Cas9 on an integrative plasmid containing...

  18. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...the proteins cooperatively can separate the DNA strands. Created with BioRender.com. Once DnaA breaks...it recruits helicases to continue unwinding the strands. This creates two replication forks that move in...elongation phase, a DNA polymerase synthesizes a new DNA strand at each replication fork. Termination sequences...act as a feedback mechanism to reduce further transcription. Replication is also regulated by the balance... concentration is controlled at the level of translation. RepZ mRNA must fold into a “pseudoknot” held...intramolecular base pairs for the protein to be translated. An antisense RNA called Inc, also encoded by...prevents the mRNA from folding and blocks RepZ translation (Asano & Mizobuchi, 1998). This system keeps ...

  19. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ... and inhibit its own transcription. As copy number increases, RepA transcription is repressed and replication...indirectly inhibit replication through inhibiting translation of replication machinery proteins, or directly... division. They don’t simply rely on chance (or random diffusion) to make this happen.  There are several...CBPs and NTPase to go around, the plasmids are randomly positioned, leading to plasmid loss (Diaz et al... if these do not provide clarity, a wet lab transformation experiment will ultimately inform whether your... Partition-associated incompatibility caused by random assortment of pure plasmid clusters. Molecular ...classification and identification based on replication and transfer systems and host taxonomy. Frontiers in Microbiology...

  20. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...fluor. Transcribing and translating a protein fluorophore also takes time - bypassing translation altogether... could be screened for their affinity to a wide range of non-nucleotide targets. These RNA molecules, ...The key to aptamers’ flexibility is their broad range of targets, including proteins, peptides, amino ...ions, and cells (even pathogenic bacteria). That range is possible because aptamers can either fold to ...biosensors. Aptabeacons Fluorescence resonance energy transfer (FRET) technology has been studied and used in...fluorescence-activated cell sorting (FACS) of E. coli transformed with aptamer libraries, the Jaffrey group selected...
Showing: 441 - 460 of 996 results