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  1. Overcoming the Challenges of Lentiviral Production

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    Blog Post
    ...elements among multiple plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid...By dividing the viral elements across multiple plasmids, the risk of recombination and consequently, the...thereby affect the titer of the virus. Additional plasmids will also affect experiment optimization. Before...different ratios of transfer, envelope, and packaging plasmids in order to obtain the best titers. Requiring ...safety by dividing the viral genome into additional plasmids thereby increasing the number of recombination...transfection and titers) To produce lentivirus, the plasmids encoding the structural elements, envelope, and...and 72h produces high titer virus for transfer plasmids with small inserts (such as CRISPR guides) while...

  2. Popular Retroviral Vectors and Their Uses in Scientific Research

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    Blog Post
    ...separate envelope and packaging plasmids as well as transfer plasmids. This concept ensures safe handling...gamma-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G, may be... genes and transgene expression across several plasmids. The most commonly used viral vector systems are...popular packaging and gamma-retroviral envelope plasmids here. Lentiviral vectors are very versatile tools...guide. Popular packaging and lentiviral envelope plasmids are listed here. Please note that both lentiviruses...provided by the Qi lab. Learn more about CRISPR plasmids for transcriptional inhibition. Retroviral Plasmid...lab barcoding library consists of 96 retroviral plasmids (based on the Murine Stem Cell Virus, MSCV) each...

  3. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

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    Blog Post
    ...Addgene.org Find plasmids from the Bethune Lab Find Plasmids from the Roux Lab Find Plasmids for FRET ...BirA* fusion proteins are expressed in cells via plasmids or by the creation of stable cell lines and these...Interested in using Split-BioID with your research? Plasmids are available from Addgene! References Schopp... Central PMCID: 5467174. Find the Split-BioID Plasmids Here! Munter, S.D., Görnemann, J., Derua, R.,...site than in Schopp et al. Find these Split-BioID Plasmids Here!   Burke, B., Kim, D.I., Roux, K.J., & Raida...PubMed Central PMCID: 3308701. Find the BioID Plasmids Here! Macneill, S., & Varnaitė, R. (2016). Meet...techniques for visualizing protein turnover Learn about plasmids for in vivo biotinylation of bacterial fusions...

  4. Addgene’s Top 10 Blog Posts from 2014

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    Blog Post
    ...Melina Fan Plasmids 101: The Promoter Region – Let's Go! By Kendall Morgan Plasmids 101: Mammalian...Joanne Kamens Plasmids 101: Antibiotic Resistance Genes By Marcy Patrick Plasmids 101: Yeast Vectors...your plasmid and cloning questions through our Plasmids 101 series and by responding to your comments....detailed information about this new technology. Our Plasmids 101 series continues to grab scientists’ attention...Genome Editing By John Doench (Guest Blogger) Plasmids 101: Origin of Replication By Kendall Morgan ...CRISPR-Cas9 FAQs Answered! By Caroline LaManna Plasmids 101: What is a plasmid? By Margo R. Monroe ...

  5. Download Science-Themed Zoom Backgrounds from Addgene

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    Blog Post
    ...lentiviral plasmids include popular envelope and packaging plasmids as well as transfer plasmids. Download... the plasmids that Addgene offers by visiting our collections page. Here, you can browse plasmids by field...cute mascots!  Blugene has been helping us make Plasmids 101 videos and needed a break so they're hanging...   Do you do a lot of transformations with the plasmids you receive from Addgene? Our bacteria background...is probably a good fit! Did you know that most plasmids from Addgene are shipped as bacterial stabs? You...

  6. DNA Purification Without a Kit

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    Blog Post
    ...absorb the nucleic acids. Containinants bind cellulose less tightly than nucleic acids, so placing the dipstick...contaminants while nucleic acids stick around. Finally, purified total nucleic acids are released by dipping...bacterial cultures or agarose gel slices Product: plasmids or DNA fragments The plasmid purification and ...mammalian cells or tissue Product: total nucleic acids While glass or silica are commonly used to bind ... efficiently binds, or at least entraps nucleic acids. The Botella lab took advantage of this property...pipette.  Here’s how this method works: nucleic acids are broken out of cells with a solution of salts...Additional resources on the Addgene blog Read our Plasmids 101 blog series Find other plasmid protocols and...

  7. Lentiviral Vector Uses and Overview

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    Blog Post
    ...system consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with...genome integration. After co-transfection of these plasmids, 293T cells released transgene-containing lentiviral...recombination between the transfer and packaging plasmids does exist. To further reduce this possibility... First, the packaging system is split into two plasmids: one encoding Rev and one encoding Gag and Pol...transgene-expressing or knockout cell lines. Many CRISPR plasmids are designed for lentiviral use, including Addgene...impact and safety. Many of Addgene’s most requested plasmids are used with lentiviral expression systems. If...

  8. A new platform for fast functional characterization of potentially cancerous mutations

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    Blog Post
    ...collection of plasmids each carrying a single gene with a defined mutation or gene fusion. Plasmids carrying...Resources on Addgene.org Find plasmids for your research Find cancer research plasmids and resources ...mutations, and the group has deposited several hundred plasmids from this study with Addgene. Read on to learn...learn more about their work, and how these plasmids can help cancer researchers studying an assortment of...cancer research community, but what else are these plasmids useful for? They could be used to more thoroughly...phenotypic effect in a different system. These plasmids can also help scientists understand how a given...

  9. To Codon Optimize or Not: That is the Question

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    Blog Post
    ... usage bias in Plasmids 101: Codon usage bias Read more about plasmids in our Plasmids 101 blog series...There are 64 different codons that encode 20 amino acids and three stop codons, meaning that the same amino...prefer certain codons over others for certain amino acids. This is termed codon usage bias. In fact, some ...transfer RNAs (tRNAs). tRNAs bind to and deliver amino acids to the ribosome where they are incorporated into...depicts the 64 codons that encodes the 20 amino acids and three stop codons. (B) Example of codon usage...sequence databases: status for the year 2000. Nucleic Acids Research 28:292–292 . https://doi.org/10.1093/nar...usage bias, and its potential applications. Nucl Acids Res 15:1281–1295 . https://doi.org/10.1093/nar/15.3.1281...

  10. What's New in CRISPR - March 2020

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    Blog Post
    ...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...series, we’ll highlight a few of the new CRISPR plasmids available at Addgene. We will still periodically... Bioengineering Find the optogenetic repressor plasmids Cytosine base editors with minimized off-target...paper in Nature Biotechnology Find the base editor plasmids CRISPRi in Caulobacter crescentus Figure...once. Read the paper in mBio Find the CRISPRi plasmids A multiplexed gRNA piggyBac transposon system... Read the paper in Scientific Reports Find the plasmids Enhanced CRISPR-based DNA demethylation ...paper in Nature Communications Find Casilio-ME plasmids If you have a new CRISPR tool you’ve recently...

  11. Five Popular Model Organisms, Part 2

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    Blog Post
    ...Model Organisms blog post Read more about plasmids in our Plasmids 101 blog series Learn more about CRISPR...collection page Find more plasmids and resources for plant resources on Plant Plasmids Collection Page Read...in plasmid manipulation and genetics research. Plasmids are ideal tools for studying genetic modification...disease models.  Check out Addgene’s collection of plasmids containing Rattus norvegicus genes! Mexican axolotl...several vectors for use in the axolotl including plasmids for use in CRISPR and Tol2 transposase induced... and after embryogenesis. Addgene has over 600 plasmids that contain Xenopus genes and a tool-kit from...Read our molecular biology reference Find plasmids for your research ...

  12. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

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    Blog Post
    ...for creating plant expression plasmids, including plant CRISPR plasmids from the Yiping Qi lab and the...plant-specific modifications to commonly used CRISPR plasmids are necessary to use the CRISPR system in plant...et al., 2013). Addgene carries Cas9-containing plasmids for knockout, activation and repression of target...systems. Luckily, you don’t have to look far for plasmids that have the plant-specific modifications required...targeting your favorite plant gene; you can find many plasmids for a wide variety of CRISPR applications in plants... available through Addgene. In addition to the plasmids described above, Addgene carries several useful...Plant Parts Kit from the Patron Lab.  As with all plasmids in the repository, we highly recommend reading...

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

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    Blog Post
    ...Michael Davidson fluorescent protein collection Check out Addgene’s fluorescent protein plasmids and resources...Biotechnology which allows researchers to deposit plasmids containing the fluorescent protein mNeonGreen....with other fluorescent proteins. Find mNeonGreen plasmids here! mTFP1: a monomeric teal fluorescent protein...physiologically relevant changes in pH. Find mTFP1 plasmids here! mWasabi: a bright green fluorescent protein...excite at 400 nm while EGFP does. Find mWasabi plasmids here! One of the most notable things about these...are best suited for certain studies. If you have plasmids containing mNeonGreen, mTFP1, or mWasabi that ...with the scientific community, please deposit the plasmids at Addgene! References Ai, Hui-wang, et al. "...

  14. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

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    Blog Post
    ...vectors (LTRs and ITRs), plasmids with multiple polyA sites, and other plasmids that contain repeating ...I help troubleshoot involves viral vectors and plasmids with repeating sequences. Repeating sequences ...other very much undergo recombination, two smaller plasmids are formed. Viral vectors are prone to recombination...and 3’ LTR regions, it splits into two smaller plasmids. The ~3.7 kb recombined vector backbone contains...elements. Knowing how to amplify recombination-prone plasmids and testing your prep can save you the headache...browse the table of common gene mutations in our Plasmids 101: Common Lab E. coli Strains blog post. Check...copy number, try following protocols for low copy plasmids. For example, using a longer growth period (up...

  15. Addgene-Seeding Labs plasmid grant round 3: microbial drug resistance and biofilms, antidiabetic plant components, visualizing brain injury - and a glowing robot fish

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    Blog Post
    ...to provide scientific resources, equipment, and plasmids to researchers in resource-limited settings. More...the Addgene-Seeding Labs plasmid grant provide plasmids to these researchers. In the third round of the...biofilms formed on various structured surfaces. Plasmids expressing fluorescent proteins will be used to...microscopy and FACS. In an additional approach, plasmids such as pACBSR-hyg and pFLP-hyg will be used to...regulation and diabetes will be expressed from plasmids (find more information on bacterial expression...information processing during this condition is crucial. Plasmids expressing fluorescent proteins and a new two-...create this bacterial sensor, he will construct plasmids containing promoter regions that activate fluorescent...

  16. GCE4All: Making Genetic Code Expansion Accessible

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    Blog Post
    ...which non-canonical amino acids (ncAAs, also sometimes called unnatural amino acids) are placed in a protein...probably quite interested in the canonical 20 amino acids. But in your quest to learn more about your protein...protein of interest, you may find the available amino acids a bit…limiting. It may be time, then, to look towards...and synthetase introduced to the bacteria using plasmids. Rapid expansion occurred, with hundreds of new...Blue Flame award to identify highly requested GCE plasmids. But they’d also love scientist interested in,...and resources, and then checking out all of the plasmids and bacterial strains (available at Addgene for... Site-specific Incorporation of Unnatural Amino Acids into Proteins. Science, 244(4901), 182–188. https...

  17. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

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    Blog Post
    ...stationary phase, the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold (for p15a, ...Researchers express genes of interest from plasmids in order to study gene function or to engineer cells...molecular biologist to do? Let’s take a look at how plasmids recently deposited by Chris Voigt’s lab help keep...gene expression and plasmid number. In a suite of plasmids modified from pSC101 with copy numbers ranging...Because pSC101 is not representative of all types of plasmids, the stabilized promoters were also tested in ..., R6K, and ColE2 plasmids) as cell division slows (Kittleson et al., 2011). Segall-Shapiro et al. showed...collections page Download the Plasmid 101 eBook See all plasmids deposited by the Voigt lab ...

  18. All in a Twist: dsRNA

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    Blog Post
    ...suffice. This can result in DNA:RNA hybrids. Unlike dsRNA, DNA:RNA hybrids are abundant in normal cells and...when using these reagents. RNA:DNA hybrids Persistent RNA:DNA hybrids that arise during transcription are...Endogenous sources of duplexed RNA DNA:RNA hybrids Sometimes ssRNA just needs a buddy, and even DNA...throughout a single cell, so these nucleic acid hybrids are always around. R-loops are particularly enriched...substrates and in distinct cell cycles. When RNA:DNA hybrids levels are allowed to reach very high or low levels...al., Human proteins that interact with RNA/DNA hybrids. Genome Res., 28(9): 1405-1414 (2018). 10.1101/...

  19. Ghanaian Researchers to Receive Plasmid Awards from Addgene and Seeding Labs

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    Blog Post
    ...construction of new plasmids. Dr. Duedu will also use polymerases produced using plasmids from Addgene to ...doctors Bengyella and Duedu and how they plan to use plasmids from Addgene to advance their research. Louis ...cellulosic biomass. Dr. Duedu would like to use the plasmids he acquires through Addgene to produce molecular...fluorescent proteins. Dr. Duedu’s plans for these plasmids have a heavy focus on teaching. He hopes to teach...in environmental samples via PCR. We hope these plasmids will help Dr. Duedu inspire new Ghanaian scientists...and creative ideas applicants will propose for plasmids from the Addgene community and hope we can do ...

  20. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

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    Blog Post
    ...separate (and thus smaller) plasmids. Finally, having four separate plasmids allows researchers to partially... four viral genes are each encoded on separate plasmids and are co-transfected into cells alongside a ... the same plasmid (as opposed to four separate plasmids), there are a few reasons arguing against this...Technical tips It is important that all of these plasmids (those containing NP, VP35, VP30, L, and the minigenome...extremely important. For example, particular ratios of plasmids encoding NP and VP35 allow for efficient polymerase... It is important to note that this system uses plasmids where the genes (either viral genes or minigenome...Resources on Addgene.org Browse Our Viral Vector Plasmids Read Our Lentiviral Vector Guide Read Our AAV ...
Showing: 221 - 240 of 878 results