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We narrowed to 801 results for: ran

Showing: 361 - 380 of 801 results

  1. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...specialized strains are available to confer greater transcriptional control, assist with proper protein folding...harboring this mutation are deficient in outer membrane protease VII, which reduces proteolysis of the...Lacks functional RNaseE which results in longer transcript half-life F- ompT lon hsdSB (rB- mB-) gal dcm...Chloramphenicol (pRARE) Good for “universal” translation; contains 7 additional tRNAs for rare codons ...pLemo (CamR) Expression of toxic, insoluble, or membrane proteins  T7 Express   IPTG-inducible expression...addition of IPTG. T7 RNAP is then available to transcribe the gene of interest from a T7 promoter on the... the expression plasmid in order to repress transcription from a hybrid promoter. Although the DE3/T7 ...

  2. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    Published
    Dec. 20, 2017, 2:33 p.m.
    ...the non-profit sector are transferrable to industry such as technology transfer, communications, medical... minimum (8 U.S. Code § 1184 - Admission of nonimmigrants). The foreign worker must possess at least a...and therefore the academic H-1B visa cannot be transferred to an industry position. If you cannot get an... facility management, collaboration management, grant administration, clinical operations, and so on. ...include evidence such as proof of US government grants that the researcher is working under in the lab...new accomplishments to add. Obtain copies and translations of documents that are not in English. Create...    Additional Resources on the Addgene Blog Transferable Skills Guide 6 Steps to Submitting a Resume ...

  3. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ...of different fluorescent protein are available (Cranfill et al., 2016 and FPbase.org). A number of variants... organelles (Figure 1), showing that its acid tolerance is maintained in cells.   GFP is big GFP is a ...specific RNA stem loop-structure (Querido and Chartrand, 2008). Finally, a nuclease-deficient Cas9 can...clearly the case when proteins undergo post-translational modification with fatty acyl chains. For instance...Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells." Science 296.5569 ...28360214. PubMed Central PMCID: PMC5385933. 6. Cranfill, Paula J., et al. "Quantitative assessment of ...PMC2644460. 12. Querido, Emmanuelle, and Pascal Chartrand. "Using fluorescent proteins to study mRNA trafficking...

  4. March for Science

    Type
    Blog Post
    Published
    April 21, 2017, 2:30 p.m.
    ...working full-time, and have received a research grant in the last five years as polled in the American...climate change page on whitehouse.gov, freezing grants, and proposing to defund aspects of research at...following subjects: Climate change Vaccines GMOs Transgender issues Reproductive health Immigration Pollution...while many also seek to honor and credit women, immigrants, and minorities who have had great impacts on...encouraged by “Broader Impacts” requirements for certain grants) (28). As one young scientist, Matthew Niederhuber...Post: Trump administration tells EPA to freeze all grants, contracts The Guardian: Trump to scrap Nasa climate... Scientists Find a Voice at Massive Rally for Immigrants National Science Foundation: Universities Report...

  5. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ...homology-directed repair (HDR) of a CRISPR/Cas9 induced double-stranded break. This site-directed genome editing requires...methylation-dependent endonuclease (i.e. DpnI), and bacteria are transformed with the nuclease-resistant nicked plasmid (the...products of the PCR reaction are pieces of double stranded, circular DNA with staggered nicks, recent data...final PCR products are pieces of linear, double stranded DNA with homologous ends. These homologous ends...the adenosine of the GATC recognition site. Transformation: After the PCR reaction, no ligation is required...required since the E. coli you transform your PCR products into will efficiently patch up the DNA. The ...parental plasmid provides a mean for selecting for transformants which have taken up the mutagenized plasmid....

  6. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    Published
    May 10, 2016, 2:30 p.m.
    ...therapeutics - one using dCas9-SAM to activate HIV-1 transcription and destroy infected cells, the other using ...histone deacetylase (HDAC) inhibitors to increase transcription of the integrated HIV-1 genome. For a targeted...dCas9-VP64 fusion commonly used to activate transcription, but with two additional activation domains ...domains (MS2 and p65) to enhance transcriptional activation. Zhang et al. designed multiple gRNAs to target ...functional Cas9 nuclease, so there’s no potential for aberrant DNA cleavage. Direct HIV-1 cleavage allows T-cells...rates of off-target cleavage. In both cases, translating the in vitro work into an animal model comes ...cleavage. Despite the potential difficulties in translating these findings into a therapy, these papers present...

  7. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    Published
    Aug. 4, 2015, 2:30 p.m.
    ...Verkhusha VV. A photoswitchable orange-to-far-red fluorescent protein, PSmOrange. Nat Methods. 2011 Jul 31;...proteins Newly developed photoconvertible FPs like PsMOrange and mEos add another layer of complexity to imaging...GFP spreads through the cytosol via cytoplasmic strands. Image from Sattarzadeh et al., Sci Rep. 2015 CC...most useful for labs that already have created a transgenic GFP-containing organism,” says Hanson, “using...unexpectedly photoconvertible GFP proteins to make a new transgenic line could avoid problems in localization that...

  8. Easing the Protein Purification Process with pCri

    Type
    Blog Post
    Published
    June 19, 2015, 3:08 p.m.
    ...contains the MISTIC (membrane-integrating sequence for translation of integral-membrane protein constructs...glycosylate proteins and may recapitulate the post-translational modifications added to proteins in other eukaryotes...includes a special feature for difficult-to-purify membrane proteins. pCri vector pCri-13a, designed for use...protein tag. MISTIC drags tagged proteins into the membrane and may improve expression of proteins that natively...

  9. What's New in CRISPR - September 2019

    Type
    Blog Post
    Published
    Sept. 10, 2019, 12:56 p.m.
    ...proteins, or transcriptional regulators. In this system, Cas9 generates a double-stranded DNA break at...are complementary to either side of the double-stranded break in the genomic loci of interest. As the ... Find the CasX plasmids Drug inducible transcriptional activation and genome editing Yu Wang's lab...developed drug inducible CRISPR/Cas9 systems for transcriptional activation and genome editing by fusing Cas9...library contains ~93,000 unique gRNAs that target random positions along the E. coli MG1655 genome with ...

  10. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    Published
    June 27, 2023, 1:15 p.m.
    ... charged ions to rapidly pass through the cell membrane and initiate an action potential - a very useful...and close when exposed to light and change the membrane potential. OptoXRs are modified GPCRs. When exposed...downstream signaling pathways that can change both the membrane potential and a host of other cellular functions...activity upon exposure to light within a specific range of wavelengths. Excitatory channels allow positively...Photoswitches are great for teasing out important branch points along a signaling cascade that may go awry...optimal wavelength to use but can be activated by a range of wavelengths. While this may give you a little...Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proceedings of the National Academy of...

  11. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    Published
    July 28, 2016, 2:30 p.m.
    ...orthogonal attB-attP pairs per recombinase may be arranged on a register (Fig. 1C below). If attB-attP pairs...approach is to interrogate state with qPCR. The re-arrangement of DNA segments in each state of a register enables...sensors do not necessarily have to be based on transcriptional regulation, as long as they can control recombinase...recombinase activity. For example, post-transcriptional recombinase activation via RNA toehold riboregulation...expression (2). Our paper (1) explains how to arrange attB-attP pairs on a register to record all permuted... of a set of chemicals. Specific examples of arrangements that should be able to record all permuted substrings... this is to prevent interference between the transcribed gene and its 5’ UTR. Various self-cleaving hammerhead...

  12. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    Published
    May 2, 2023, 1 p.m.
    ... gene, enhancer, and promoter – all rely on the random integration of a reporter construct in close proximity...integrate within an exon of a gene to generate a transcriptional fusion. This may disrupt the endogenous function...spliced into the gene product resulting in a transcriptional fusion, much like promoter traps. While this...likely disrupt the gene it monitors due to transcriptional fusion.   Examples of promoter, enhancer...monitored genes and mutants - viral integration, transposon, and many more! Check out Addgene’s catalog to...resources on the Addgene blog Plasmids 101: Using Transposons in the Lab Plasmids 101: Screens vs. Selections...

  13. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    Published
    May 21, 2020, 1:15 p.m.
    ...visualize individual/non-overlapping cells. While transgenic animals are commonly used to specifically manipulate... to trigger recombination and expression of a transgene (Lin et al., 2018). However, this system requires...tools in Drosophila (Fig. 1):  GAL4-UAS, a transcription factor binding site. GAL4 expressed in the fly...site to cleave and inactivate any read-through transcripts of the effector gene. Effector genes, any gene...across cell types and organisms by replacing the transcription factor binding site and effector in the construct... the SPARC system to use other experimental transcription activation systems, such as LexA/p65 or mCD8...

  14. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    Published
    Aug. 19, 2014, 3:39 p.m.
    ...). If you can get away with generating transiently transfected cells, you will not need a mammalian selection...mammalian selection cassette to screen for positive transfected cells. The addition of an in-frame fluorescent...create a new strain with a disrupted gene by transforming wild-type yeast with an integrating vector expressing...majority of fly vectors are based on p-element, random or targeted, integration into the fly genome and...applications used to study gene function. From making transgenic animals, gene targeting and genome modifications...

  15. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...positively transfected. Alternatively, you can use a selection marker to select transfected cells (for...nickase doesn't have the ability to induce double-stranded breaks like the wildtype Cas9 does. There are ... The human U6 promoter prefers a 'G' at the transcription start site to have high expression, so adding...Before starting your experiment, consider co-transfecting with GFP. This allows you to sort for GFP-positive...resistance cassette, such as PX459). After you co-transfect the CRISPR/Cas system with your homologous recombination...

  16. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    Published
    April 25, 2019, 3:01 p.m.
    ...protein (LanYFP) from the marine invertebrate Branchiostoma lanceolatum in 2012. Using structural modeling...mNeonGreen can be used as a fusion tag for a wide range of applications from traditional imaging to FRET...Alberta and University of Oregon created mTFP1 by transforming E. coli with a gene library of approximately...mTFP1 can be used as a FRET donor to mCitrine and mOrange or for live cell imaging of fusions and FRET reporters...particularly with fluorophores that excite in the violet range. For example, mWasabi can be used with Sapphire ...monomeric green fluorescent protein derived from Branchiostoma lanceolatum." Nature methods 10.5 (2013): 407...

  17. Addgene Begins Distribution of Recombinant Antibodies

    Type
    Blog Post
    Published
    March 28, 2022, 4:30 p.m.
    ... our values of transparency, reproducibility, and community-building. The transparency begins with sharing...Addgene’s antibody resource is funded by a generous grant from the NIH’s Brain Research Through Advancing ...share plasmids with each other. Together, we can transform the antibody sharing landscape as well.     References... Improving Reproducibility through Rigor and Transparency of Experimental Methods. Neuron 90(3):434-6.... project is supported by a NIH BRAIN Initiative grant (U24NS119916 ). The content of this article is solely...

  18. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

    Type
    Blog Post
    Published
    Dec. 13, 2018, 1:14 p.m.
    ... AAV2 interacts with the packaging cell's cell membrane When packaging adeno-associated viral vectors ...to be due to the viral particle’s affinity for heparan sulfate proteoglycans on the surface of the packaging...supernatant, the viral particles associate with the cell membrane and are taken into the cell via endocytosis. In...looking to improve their AAV yields. Given the wide range of downstream applications scientists rely on our...preparation. Through sugar detection assays, in vitro transduction, and silver stain approaches we show that there...test the endocytosis of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe...

  19. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    Published
    March 7, 2023, 2:15 p.m.
    ... method entails arranging all calculated p values from smallest to largest and ranking them (1 being the...dispersion – standard deviation/error, variance, data ranges. how closely data points fall to each other (see...significance by finding the largest p value where its rank divided by the total number of comparisons performed...– Tukey’s test, Newman-Keuls, Duncan’s multiple range test, and several others. Each test has different...negatives, moderate false positives. Duncan’s multiple range test – Most powerful. Low false negatives, high ...): 126-130 (2012). 10.1097/SMJ.0b013e3182498ad5 Curran-Everett, Douglas. Explorations in statistics: statistical...

  20. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    Published
    Oct. 9, 2018, 12:21 p.m.
    ...postdoctoral fellows (PFs) face a number of challenges ranging from long periods of training to limited job opportunities...preparation) A history of successful funding (fellowship grants) A project with sufficient preliminary data to...your own lab and get an independent investigator grant (e.g. an R01 from the NIH, in the United States)...postdoctoral mentor. A good mentor typically writes the grants that fund your research, guides the research directions... available for mentees due to travel, teaching, grant writing, etc. Also, it is unlikely that a mentor...reference, informal recommendations, more positive grant application and manuscript reviews, etc. You'll ... Non-Academic Career Decisions Check out our transferable skills guide Resources on Addgene.org Learn...
Showing: 361 - 380 of 801 results