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We narrowed to 1,038 results for: LAS

Showing: 301 - 320 of 1038 results

  1. Degrading DNA with Cascade-Cas3

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    Blog Post
    ...Cascade-Cas3 plasmids in Addgene's collection! A “complex” system Cas3 is part of the Class 1 CRISPR family...complexity. Figure 1: Schematic of Class 1 CRISPR types. Class 1 includes type I with a Cas3 effector...out the brunt of the mechanical work for editing. Class 1 CRISPR systems make up the majority of CRISPR-...CRISPR-based immune systems in bacteria and archaea. Class 1 is further broken down into different types, with...making up type I, the most abundant type in the Class 1 category. These type Is can then be further broken... Various labs have designed all-in-one Cascade plasmids — such as the Tomoji Mashimo lab and the Tilmann...Koonin, E. V. (2015). An updated evolutionary classification of CRISPR–Cas systems. Nature Reviews Microbiology...

  2. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...packaging plasmids: the pHelper plasmid containing Ad E2A, E4, and VA RNA genes; and the pRepCap plasmid containing...with your transfer plasmid, carrying your gene of interest, and the two helper plasmids, commonly called...blog Parts of the AAV Packaging PlasmidParts of the AAV Transfer PlasmidAAV Variables That Matter AAV protocols...called “triple plasmid transfection” (Figure 1).    Figure 1: Overview of key steps of AAV production, iodixanol...repeats (ITR). You should always have a high-quality plasmid standard and rAAV reference material of a known...before undergoing titering, to remove residual plasmid DNA carried over from the production process. For...

  3. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

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    Blog Post
    ...Browse our CRISPR interference plasmids Find bacterial CRISPR plasmids Read our CRISPR guide ...transposon system. This system has two parts: a plasmid expressing the Tn7 transposition genes (tnsA, tnsB... tnsB, tnsC, tnsD, and tnsE), and a plasmid that contains the Mobile CRISPRi cassette flanked by left ...copy of the RP4 transfer machinery mobilize these plasmids for transfer to recipient cells during a tri-parental...Mobile-CRISPRi ICE element forms a conjugative plasmid-like DNA circle and is transferred to recipient...transposon containing the CRISPRi components and a plasmid with the Tn7 transposition genes are transferred...RP4 transfer machinery, which mobilizes the Tn7 plasmids, and lack the ability to synthesize diaminopimelic...

  4. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

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    Blog Post
    ...Fusions Plasmids 101: Secondary Nanobody Toolbox Resources on Addgene.org View the Antibody plasmid collection...The Seeger lab found a way around this by using plasmids to generate sybodies, synthetic nanobodies against...nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target epitopes that...form of mRNA for academic research. The template plasmids for generating your own libraries are also available...available in the Sybody Generation Toolbox plasmid kit: SB_concave, SB_loop, SB_convex. These scaffolds...efficiently subcloned into the phagemid pDX_init plasmid, using the fragment exchange (FX) cloning technique... subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for the production of tag-free proteins for further...

  5. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...lentiCas9-blast. A549 cells were transduced (MOI = 37) and selected with 1 µg/mL blasticidin for 9 days...18007609. Addgene Materials Used 1. lentiCas9-Blast (Plasmid #52962) was a gift from Feng Zhang and is described...BRDN0000561167) (Plasmid #80034) and BRAF gRNAs (BRDN0001145345 and BRDN0001146266) (Plasmid #76479 and #76480...transduction with Cas9 lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells were then used ...expression in A549 cells transduced with lentiCas9-blast. Anti-Cas9 Western blotting was performed on whole-cell...then expanded for an additional 3 weeks under blasticidin selection. Anti-Cas9 Western blotting was performed...viruses, but you can find a list of validated gRNA plasmids on our website. There are a few reasons to target...

  6. Bringing Sustainable Practices to the Lab: Innovations

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    Blog Post
    ...labels! Reusing plastic Historically, scientists washed and reused materials such as glass pipettes or inoculation...that uses this waste as plastic lumber for things like truck bumpers, plastic adirondack chairs, and synthetic...other scientists save resources by sharing plasmids Find plasmids at Addgene ... popular theme in life sciences, some of these plastics are high grade and can be washed and reused. This...tips cost in half, not to mention the amount of plastic you’ve saved. Repurposing regulated medical waste...use this material to produce various grades of plastic lumber and distribute it to industrial and residential...

  7. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ...is mediated by ADAR2DD, not PspCas13b. Find the plasmids for the RESCUE RNA editors here! Testing and improving...for viral delivery.  Find the RESCUE RNA editing plasmids here! Future directions Building on previous RNA...Science (2017):pii: eaaq0180. PMID: 29070703. Find plasmids from this paper at Addgene. Abudayyeh, Omar O...550(7675) (2017):280-284. PMID: 28976959. Find plasmids from this paper at Addgene. Abudayyeh, Omar O...27256883. PubMed Central PMCID: PMC5127784. Find plasmids from this paper at Addgene. East-Seletsky, Alexandra...) (2016):270-273. PubMed PMID: 27669025. Find plasmids from this paper at Addgene. Gootenberg, Jonathan...28408723. PubMed Central PMCID: PMC5526198. Find plasmids from this paper at Addgene. East-Seletsky, Alexandra...

  8. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    ...stability of modified Ti plasmids during plant infection, as well as plasmid yield during preparation ...GreenGate Plasmids Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More Plasmids 101: Origin...Agrobacterium carries a “tumor-inducing” or Ti plasmid, which allows it to transfer genetic material into...in two species: Agrobacterium and E. coli. Like plasmids used in other systems, binary vectors are cloned...It’s a lot of information to fit into a single plasmid, so most first-generation binary vectors are quite...pSiM24 are suitable for many applications, the plasmid can also be customized due to its modular structure...Addgene Check out Addgene’s plant vectors! Find the plasmids from this paper at Addgene. Other relevant blog...

  9. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...protein in question: C2c1, C2c2 and C2c3 (short for Class 2, candidate 1, 2, or 3). C2c1 and C2c3 are related...to see this group of enzymes continue to make a splash in the CRISPR field. We feel that the culture of...27256883. PubMed Central PMCID: PMC5127784. Find plasmids from this paper at Addgene. 2. East-Seletsky,...) (2016):270-273. PubMed PMID: 27669025. Find plasmids from this paper at Addgene.  3. Gootenberg, Jonathan...28408723. PubMed Central PMCID: PMC5526198. Find plasmids from this paper at Addgene. 4. East-Seletsky,...) (2017):373-83. PubMed PMID: 28475872.  Find plasmids from this paper at Addgene.  5. Abudayyeh, Omar...550(7675) (2017):280-284. PMID: 28976959. Find plasmids from this paper at Addgene. 6. Shmakov, Sergey...

  10. Tips for Improving Your Next Manuscript

    Type
    Blog Post
    ... unsure if I should possibly take a composition class in the English Department. He suggested that I sign...writing. Following his advice, I applied to the class. Receiving the email that I had been accepted into...Cotter, Mary O’Riordan, and Victor Di Rita, and, last, homework assignments. The instructors are leaders...microbiology with multiple high-profile publications. My classmates consisted of graduate students, post-grads, and...early career scientists. Over the course of the class, the moderator discussed individual sections of ...The assignments, which consisted of reading that class’s section of a manuscript and critiquing it, were...was time to work! Set up similarly to the online class, we met as a group for sessions covering individual...

  11. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    ....12137. PubMed. Find the Plasmids at Addgene Kowarz Lab plasmids Izsvak Lab plasmids  ...the gene of interest (blue) in a plasmid backbone. A separate plasmid contains the transposase gene (red...). The original plasmid is empty following the removal of the transposon; the plasmid is then degraded...16 variants with different selection markers (blasticidin, neomycin, puromycin, hygromycin) and fluorescent...for insertion in the correct orientation. These plasmids use a novel synthetic RPBSA promoter, which is...

  12. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

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    Blog Post
    .... pBLADEONLY: This plasmid contains only BLADE and is based on the pTRC99a plasmid. It does not include...AraC and other promoter systems, head over to our Plasmids 101: Inducible Promoters article.) Modifying AraC...reporter downstream of the PBAD promoter. To use this plasmid with your gene of interest, you would replace mCherry...PBAD promoter. If you already have a pBAD33-based plasmid expressing your gene of interest, all you have ... and shared with Addgene. You can find all the plasmids deposited from Di Ventura's lab, including the...nuclear export system (LEXY) here. Optogenetic plasmids from the Khammash lab can also be found here. ...Learn more about inducible promoters Visit the Plasmids topic overview page Resources on Addgene.org ...

  13. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ...the cytometer illuminates the passing cell with a laser and detects the light scattered by that cell at ...surface. Bear in mind, the voltage setting of the laser impacts the magnitude of FSC, SSC, and all fluorescent...experienced colleagues.   Figure 1: The laser pulse illuminates the cell and is scattered forward...the time that the cell spent passing through the laser beam. A is simply the area under the resulting curve... Figure 2: As the cell passes through the laser beam and causes it to scatter (FSC/SSC), the detected...), the time the cell needed to pass through the laser (W) and the area under the curve (A).   While... FSC/SSC plot, largely depends on cell type and laser voltage. The voltage describes an electric potential...

  14. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    ...Addgene.org Find Fluorescent Protein Plasmids Check Out Other Hot Plasmid Technologies ...proteins and is driven by an inducible promoter (Plasmid 64542 or 64543) Before translation, both GFP and...contains a CAAX sequence to bind the mRNA to the plasma membrane; this sequence prevents diffusion of the...Since the SunTag-based system tethers mRNAs to the plasma membrane, it’s not appropriate for use with mRNAs... physiological states. References 1. Ingolia, Nicholas T., et al. “Genome-wide analysis in vivo of translation...19213877 PMCID: PMC2746483 Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation...1367-71. PMID: 25792328 PMCID: PMC4451088 Find plasmids from this paper at Addgene. 3. Yan, Xiaowei, ...

  15. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ...different plasmids. Harvard’s DRSC/TRiP Functional Genomics Resource Center has various plasmids for overexpression...gene of interest is cloned into a plasmid, you can send your plasmid to a company. For a few hundred dollars...Basically, you place your gene of interest in a plasmid and get the vector incorporated into the germline...filtering by fly as species of gene. Find Gal4 and UAS plasmids at Addgene Choosing a vector to generate a transgenic... dollars, they will inject the plasmid into fly embryos and pick out flies where your transgene has been...incorporated into the germline. Outsourcing the plasmid injections is much easier than doing it yourself...Recombination Resources at Addgene.org Browse Drosophila Plasmids ...

  16. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    ...effector nuclease (TALEN) kit consisting of 834 plasmids. The researchers showed they could use their TALEN...particular DNA sequence? Musunuru: The kit includes 832 plasmids containing prefabricated pieces that allow you...the genome in rapid fashion. The two remaining plasmids in the kit have the rest of the protein in place...fluorescent proteins used to sort the cells after the plasmids have been introduced by transfection or electroporation...protein that binds those 15 base pairs is take four plasmids out of the library in the appropriate combination... ligation with the four inserts and the master plasmid in one step. The kit also has other unique aspects...you are designing a TALEN on paper to a finished plasmid. With our protocol, it is feasible to produce genetically...

  17. dTAG - You're it!

    Type
    Blog Post
    ...1). Plasmids: We generated a series of plasmids that are available from Addgene. These plasmids enable...Resources on Addgene.org Browse all CRISPR plasmids CRISPR plasmids for protein tagging Find validated gRNAs...Detailed protocols describing the use of these plasmids are available here. The Methods section of Nabet...cloning procedures used to construct the deposited plasmids. Molecules: We synthesized dTAG molecules by conjugating...terminus should be tagged, the dTAG lentiviral plasmids can serve as a rapid means of comparing functionality...Cas9 Nickase Design and Homology Directed Repair Plasmids for endogenous gene tagging in human cells Resources...

  18. Working with Nuclear Receptors

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    Blog Post
    ...problem of sharing plasmids, providing researchers with a one-stop shop for plasmids and vectors, and making...other tissues. At the Nuclear Receptor Signaling Atlas (NURSA), one of our primary goals is to promote ...progress that has been made in the NR field over the last thirty years or so would not have been possible ...development of a vast number expression and reporter plasmids and the free exchange of these vectors between...increased the burden associated with identifying plasmids in the research literature and acquiring them ... feature direct links to listings of available plasmids for our curated NRs and coregulators. We look ...edits the website of the Nuclear Receptor Signaling Atlas (NURSA) at www.nursa.org. Bert O’Malley, M.D.,...

  19. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...analyzing the plasmid and viral DNA sequences and expression profile at multiple steps from plasmid amplification...to flip back into the inverse orientation (See Plasmids 101: FLEx Vectors). While this strategy greatly...recombination that are typically employed during viral plasmid amplification (Fischer, KB. 2019). This means that...of this finding, we now include guidance on the plasmid page of DIO/FLEx AAV vectors where this occurs ... mechanism of spontaneous reversion during AAV plasmid amplification. On the left homologous pairs of ...specific” promoter, such as the truncated tyrosine hydroxylase (TH) promoter for catecholamine neurons or glial.../FLEx vectors for off-target expression.  Some classes of neurons transiently express TH during development...

  20. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

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    Blog Post
    ...Allen Institute for Cell Science Plasmid Collection Check Out Plasmids for FRET Check Out the Fluorescent..., most secreted proteins are produced in the endoplasmid reticulum, modified in the golgi and then transported... on the gene names in the figure below to find plasmids containing that particular gene. ... For subcellular markers in yeast see our plasmid collection from Sue Jaspersen (Stowers Institute...Prescott (Monash University). You can find more plasmids for labeling your subcellular structure of interest...interest in the Allen Institute for Cell Science Plasmid Collection. You can also use the library of full-length...Localization Poster here! References 1. Allen Institute Plasmid Pages 2. Cooper GM. 2000. The Cell: A Molecular...
Showing: 301 - 320 of 1038 results