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We narrowed to 62 results for: Free Use GFP

Showing: 1 - 20 of 62 results

  1. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    ...find and use the plasmids you need. Here's what you'll find in this article: An improved cell-free system... The CRISPR Corner New ready-to-use viral preps   Improved cell-free transcription/translation system...successfully reconstituted in this cell-free system. In summary, this cell free system allows for a relatively ...vectors and flow cytometry is used to sort populations by mCherry and GFP expression to check for ABE’s...examine biosynthetic pathways  By Gabrielle Clouse Recently, cell-free transcription/translation systems (also...that produce useful natural materials, such as antibiotics. Simon Moore and Paul Freemont’s lab previously...Graphical Abstract: Schematic representation of cell-free system, capable of producing fluorescent proteins...

  2. Viral Production

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    Collection
    ... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...Endotoxin contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize...addition, plasmids are typically prepared using endotoxin-free plasmid purification kits. Resources Browse... titers obtained using ddPCR are generally 2–3 fold higher than those achieved using the standard qPCR...Resources Overview of Viral Production Viruses are generated using standard methods that have been optimized...of unexpected DNA contaminants. The data is also used to determine recombination rates in FLEX/DIO/fDIO...protein present in the sample is also determined and used to determine purity of the AAV preparation. Figure...

  3. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...Gagoski used SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression...can be used, and how they’ve been used to date . If you have any questions, please write to us in the ...N-terminal eGFP tag carried on all constructs. The Cell-Free Protein Expression Kit has been used to produced...sequences used in this so-called bicistronic design (BCD). The 49 vectors in this kit can be used to drive...protein of interest. The plasmids used to generate these fusions all use the same restriction enzymes for...Purification Process with pCri Cell-free protein expression kit Cell-free expression systems have been improved...found at the 3’ end of the leader sequence. Using a variety of GFP and RFP fusions as their test genes, the...

  4. Hot Plasmids - May 2022

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    Blog Post
    ...2021.    BLITZ uses the biotin ligase TurboID fused to a conditionally stabilized GFP-nanobody rather...line expressing a GFP-tagged version of a protein of interest. The binding of the GFP nanobody to its target... proximity to the GFP-tagged protein (albeit slightly offset due to the nanobody-GFP interaction). The...be deconvoluted using freeware.    Fig. 4: Images of 12 visual barcodes in use in A375 cell line...Fig. 3: Development of a cleavage-free dCas9-based gene editor using microbial SSAPs. Image from Wang,... hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like to write...recently created a flexible system that makes use of pre-existing transgenic lines, termed BLITZ (Biotin...

  5. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    ...Toolbox: Free Tools for Making Scientific Graphics by Beth Kenkel Find a round-up of tools you can use for...scientific graphics. PS: they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right...you’d like to see more of? Let us know by leaving a comment or emailing us at [email protected].  Thank you...toxin-antitoxin systems and sacB counter-selection) tools used in cloning. 8. Bright Monomeric Fluorescent Proteins...people are using for their research. In this blog post, we’ve covered five model organisms: mouse, fruit ...Joachim Goedhart shares some strategies for generating GFP fusion proteins. From considering protein size and... some of these top blog posts. We’d also like to use this time to hear some feedback from the community...

  6. Worm Expression Resources

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    Collection
    ...available at Addgene for use in worms. Browse Plasmids Plasmid Tools for Use in Worms The table below ... a CRISPR-Cas9 system. Cloning-free CRISPR for C. elegans , which uses purified Cas9, such as that expressed... repository? Help us widen our resources for the community by depositing your useful tools , or by suggesting...as CRISPR and synthetic biology collections) for use in C. elegan research. Plasmid...Caenorhabditis elegans ( C. elegans or “the worm”) is widely used as a genetic model organism due to their rapid life...genes by feeding worms bacteria expressing dsRNA or use CRISPR/Cas9 gene editing to easily engineer specific...below highlights plasmids that contain tools for use in worm research. This includes CRISPR, RNAi, empty backbones...

  7. Viral Production at Addgene

    Type
    Blog Post
    ... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...Endotoxin contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize...addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.     Figure...titers obtained using ddPCR are generally two to three fold higher than those achieved using the standard...AAV preps and our lentiviral preps.   Viruses are generated using standard methods that have been optimized...of unexpected DNA contaminants. The data is also used to determine recombination rates in FLEX/DIO/fDIO...protein present in the sample is also determined and used to determine purity of the AAV preparation. ...

  8. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ... the GFP-Puro will be inserted just upstream of a stop codon. One potential concern is if the GFP-Puro...Integration into Target Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created ...template with flanking sequence homology for error-free DSB repair. NHEJ joins the ends of a DSB in an error-prone...insertions and deletions common. In contrast, MMEJ uses regions with 5-25 bp of microhomology flanking a...inefficiency of HDR for knock-ins, some labs have used NHEJ for whole plasmid integration; however, since...detailed protocol for MMEJ-mediated knock-in of a GFP-Puro cassette into a given locus, just upstream of...microhomology to the insertion locus flanking the GFP-Puro cassette. Three double stranded breaks are necessary...

  9. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...antibodies against them, fused them to GFP, and compared them to popularly used epitopes tags such as HA...to complement the use of optogenetic tools, where light used for excitation can cause unintended activation...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...plasmids for use in high-throughput screening of pre-mRNA splicing. This reporter system can be used for identifying... that can be used for monitoring cellular dynamics. These fusion proteins have been used as subcellular...Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B Mitochondria mito-BFP...GTPase in division) GFP-Mff (outer membrane protein) Late Endosome mCherry-Rab7A GFP-Rab7A     ...

  10. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

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    Blog Post
    ...nanobodies described here can be used to target any protein fused with GFP, YFP, mCerulean, or other related...for quantitative analysis using different detection methods. Since the anti-GFP nanobody is able to bind...a set of plasmids each containing an anti-GFP nanobody fused with a different functional element (tyrosine...University of Massachusetts Medical School have developed a group of plasmids (here) that can be used to label... hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like to write...facilitating techniques that are dependent on long, error-free, reverse transcription of RNA such as long-read ...that associate with specific genomic loci. He made use of catalytically inactive dCas9 by fusing it to an...

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

    Type
    Blog Post
    ...editing. A virus encoding GFP plus a gRNA targeting GFP (pXPR_011, 59702-LV) was used to assay Cas9 activity...from BRAF-edited and GFP-edited amplification products (BRAF-GFP duplex) and from GFP-edited amplification... 1. Use multiple gRNAs to target different regions of your gene. I used existing BRAF gRNA viruses, but...I thought I could see GFP expression with a fluorescence microscope, but the GFP expression was so low...expressed GFP) that I couldn’t get a good image. You could probably also do Western blotting against GFP expression...76480-LV) or control GFP gRNA (BRDN0000561167) (80034-LV) were individually used to transduced stable ...product indicates a gene edit. GFP gRNA-transduced Cas9-A549 cells are used as a control. DNA was analyzed...

  12. Negative Can Be Positive: Open AAV Data with Addgene

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    Blog Post
    ...literature, scientists can use the Data Hub to compare different methods and vectors, and use this data to guide...when others have already answered this question and used it to publish!  Working with AAV vectors in neuroscience... I started working with AAV vectors. I wanted to use an AAV vector to express ChR2(H134R) in A2 norepinephrine... cassette with an ubiquitous promoter to express GFP in a group of young rats. We found that, in this ...important factors such as what is the best promoter to use, or if and how much the spread of the virus to adjacent...label norepinephrine (NE) expressing neurons and GFP (green) to label neurons transduced by each serotype... (3,5,8), injection titers (1,2), and cell types used for packaging viral particles (6), can influence...

  13. Rinehart Lab Phosphoprotein Reagents

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    Collection
    ...68299 GFP E17TAG/Q157TAG Plasmid 68298 GFP S2TAG/E17TAG Plasmid 68297 GFP Q157TAG Plasmid 68296 GFP S2TAG... production of recombinant phosphoproteins using cell-free protein synthesis . Nature Communications ,...generated with either phosphoserine using SepOTSλ (Plasmid #68292) or serine using supD tRNA ( SupD (Plasmid #68307...The Rinehart Lab has developed several tools useful for the incorporation of phosphoserine into your ...characteristics, and an overall improved ease of use when expressing recombinant phosphoproteins. These...RSF1030 origins or a low copy p15a origins should be used. Find further details in Mohler et al. (2023) and...library (Pooled Library #111704) is anticipated to be useful for laboratories performing phosphoproteomic experiments...

  14. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes collected...many future variations using nanobodies or scFvs for other targets or could be used for applications beyond...that this method of delivery could be used to partially correct mouse models of genetic blindness (An et ...epigenetic silencing, which can be toxic, CHARM uses a cofactor fused to a histone tail to recruit and stimulate...In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study) are captured...SpyTag-SpyCatcher linkers and spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines ...) 3D model of the obtained structure of the H1.8-GFP-bound nucleosome. Images adapted from Arimura et ...

  15. Delivery Methods for Generating iPSCs

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    Blog Post
    ...minicircle RNA Delivery DNA-Free High High Transgene-free and vector-free Multiple transfections required...required Protein Delivery DNA-Free Low High Transgene-free and vector-free. Slow and inefficient Difficult...Excisible Medium Medium Generates transgene-free and vector-free cells Genome integration Need to sequence...Non-integrating Low Medium Generates transgene-free and vector-free cells No genomic integration Inefficient...Non-integrating Medium Medium Generates transgene-free and vector-free cells No genomic integration Can be difficult...usually Medium Medium Generates transgene-free and vector-free cells Limited genomic integration Multiple...Non-integrating Medium Medium Generates transgene-free and vector-free cells No genomic integration Inefficient...

  16. CRISPR Plasmids - Tagging

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    Collection
    ...developed a CRISPR tagging technique for use in Drosophila cells that uses PCR to generate both an expression...marker-free mutations, insertions, and deletions at any locus in C. elegans . This system uses a 10-day...proteins by using donor constructs that harbor synthetic short homology arms. Notably, they have used Auxin-...information about HDR/repair template design. How to use CRISPR to tag your gene of interest Mendenhall and...Zhang lab's PX458 . Multiple gRNA plasmids may be used. The HDR (homology directed repair) donor plasmid...cell lines. To repeat the tagging in your system, use the plasmids as listed in each row. If your own gene... and right homology arms. More details on how to use this plasmid in your experiments can be found below...

  17. Fluorescent Protein Guide: Empty Backbones

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    Collection
    ...Brightness pKa Maturation Structure Plasmids Free Use GFP (fuGFP) 395 503 Monomer pUS252 - Mammalian Expression...Expression cfSGFP2 493 517 34 5.4 Monomer (A206K) cfSGFP2-N - Mammalian Expression (cysteine-free SGFP2) ZsGreen...Mammalian Expression sfGFP (superfolder GFP) 485 507 54 14 min Prone to dimerization sfGFP-N1 - Mammalian Expression...Expression EGFP 488 507 34 6 25 min Prone to dimerization pcDNA3-EGFP - Mammalian Expression pCAG-GFP - Mammalian...Expression PA-GFP 504 517 400 14 Monomer (A206K) pPAGFP-N1 - Mammalian Expression pPAGFP-C1 - Mammalian... pRSETA-PAGFP - Bacterial Expression pFA6a-PA-GFP-kanMX6 - Yeast Expression pFA6a-link-yEPAGFP-CaUra3 ...high concentration. Proteins that are derived from GFP and contain the A206K mutation are monomeric at all...

  18. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

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    Blog Post
    ...researcher can cause them to interact by stimulation with blue light.  Similarly, light can also be used to control... hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like to write...Institute for Basic Science (IBS) has developed another useful optogenetic tool that enables robust and efficient...without the extra peptide. CRY2clust should prove a useful tool to further probe cellular function. The CRY2clust...segment Optogenetic stimulation is a powerful tool used by neuroscientists to interrogate neuronal networks...plasmids described in this work including the soCoChR-GFP plasmid as well as several derivatives encoding alternative...podcast segment With the help of engineered microbes, used coffee grounds could be economically repurposed ...

  19. Antibodies 101: Single Chain Fragment Variables (scFvs)

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    Blog Post
    ... are scFvs used for? scFvs are used in many of the ways antibodies are. scFvs are even used to screen ...GCN4 binding scFv that’s fused to GFP. When the multiple copies of the scFv-GFP fusion bind the SunTag ...clearance, which is useful for imaging applications; better tissue penetration which is useful for therapeutic...immunized animals, but it's possible to avoid animal use by using semi-synthetic or entirely synthetic VL and ...contains an HA-binding scFv that’s fused to a fluorescent protein such as GFP. The HA frankenbody works just... this, one or more of these approaches are often used for scFv expression in bacteria:  targeting to ...Sandomenico et al., 2020). conversion to a cysteine free format that’s easier to express in the cytoplasm...

  20. Fluorescent Proteins 101: When GFP lets you down

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    Blog Post
    ... There are numerous GFP variants. Here I use the name ‘GFP’, which refers to mEGFP. This variant was made...2013; Hilaire et al., 2017). GFP can only be used for tagging proteins GFP fusion proteins are made by ...S65T) in the original jellyfish GFP (AvGFP) to create enhanced GFP (EGFP), a variant with improved brightness...Microscopy (University of Amsterdam). GFP is the most popular, most widely used genetically encoded fluorescent...examined on a case-by-case basis. GFP is sensitive to acid The chromophore of GFP can exist in a protonated and...its acid tolerance is maintained in cells.   GFP is big GFP is a 28 kDa protein that resembles a cylinder... a fusion with GFP allows for detection of genomic locations (Chen et al., 2013). GFP fluorescence overlaps...
Showing: 1 - 20 of 62 results