Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 97 results for: plasmid transfection

Showing: 41 - 80 of 97 results

  1. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ...consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated...and host genome integration. After co-transfection of these plasmids, 293T cells released transgene-containing... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with...Many CRISPR plasmids are designed for lentiviral use, including Addgene’s top requested plasmid in 2015: ...itself due to a few missing proteins. The envelope plasmid contained a viral envelope to dictate the tropism...the vector could infect. Finally, the transfer plasmid encoded the desired transgene flanked by HIV-1 ...recombination between the transfer and packaging plasmids does exist. To further reduce this possibility...

  2. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    Published
    Nov. 5, 2024, 8:32 p.m.
    ...three plasmids — a transfer plasmid containing your gene of interest plus two packaging plasmids (three...baculovirus plasmid. Production of baculovirus requires only two plasmids — your transfer plasmid and the ...three packaging plasmids for 3rd generation lentivirus production). The packaging plasmids are widely available...to worry about cloning your transfer plasmid. The transfer plasmids have different packaging capacities...in mammalian cells is relatively quick. After transfection, HEK-293T cells start producing virus within...systems, it is standard to do only one passage (transfection). This differs from typical insect cell preparations...called a bacmid. Like AAV and lentiviral packaging plasmids, bacmids are readily available, so the amount ...

  3. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ...Pennsylvania Vector Core. Transfections are performed using the transfer plasmid, a plasmid encoding rep and serotype-specific...with primers targeting the transfer plasmid used in the transfection. PCR products are analyzed by gel .... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ... serotype-specific cap genes, and a plasmid encoding adenoviral helper sequences. AAV preparations are...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...offline and discarded. Confirmation of the transfer plasmid Addgene uses a rigorous barcode matching system...microbial contamination. Purity When possible, all plasmids used for viral production are propagated in the...

  4. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...approach often involves the transient transfection of expression plasmids in a given cell line. For flow cytometry...interference (RNAi). The most common method is transfection of short interfering RNA (siRNA). Knockdown ... (tagged or untagged) protein When expression plasmids are available, or the necessary expertise to clone...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 ...

  5. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ...can be produced through a single or dual-plasmid transfection, depending on whether the heavy and light...contained to one plasmid or if they’ve been separated onto two. You’ll first transfect your cells with ... great collection of ready-to-use rAbs and rAb plasmids!). Since recombinant antibodies conveniently allow...with your plasmid(s), and then wait 7-14 days (in mammalian cells) to collect the cell culture supernatant...on the Addgene blog Introduction to Antibodies Plasmid-based Recombinant Antibodies Polyclonal Antibodies...

  6. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ... target plasmid and pX330-sgRNA plasmid (Mashiko et al. 2013). The pCAG-EGxxFP target plasmid contains... (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences ... simple matter of transfecting HEK293T cells with your modified pCAG-EGXXFP plasmid along with the individual...individual gRNAs in their PX330 plasmids (or similar plasmids for gRNA expression), one gRNA at a time.... system floating around, and many pre-designed plasmids containing these variants ready for purchase. ...multi-cloning site (MCS) between EGFP fragments. The pX330 plasmid contains humanized Cas9 expression cassette and... you can insert this region into a pCAG-EGXXFP plasmid using standard cloning techniques (The XX indicates...

  7. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    Published
    Dec. 14, 2017, 2:08 p.m.
    ...maintained DNA plasmids that express reprogramming factors To date, the mRNA transfection method has not...and to model diseases in vitro. Find stem cell plasmids at Addgene Many sources of adult somatic cells...generating hiPSCs for clinical purposes: mRNA transfection- this allows transient expression of the reprogramming...group is an optimized mixture of three episomal plasmids (Table 1) encoding the reprogramming factors. ...enhance the reprogramming efficiency, an extra plasmid, pCXWB-EBNA-1 is used in the reprogramming cocktail...increases protein expression from the other episomal plasmids. When used on PBMCs, the pCXLE episomal vector...available at Addgene. Table 1: List of pCXLE episomal plasmids, their Addgene ID numbers and the reprogramming...

  8. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    Published
    Aug. 27, 2020, 1:15 p.m.
    ...This is straightforward in easy-to-transfect cell lines where plasmid dosage can be tightly controlled....because it relies on a single BiFC pair. Find BiFC plasmids for your experiment at Addgene! How to Design ...BiFC constructs are expressed transiently by transfection or stably impacts the success of BiFC experiments...Resources on Addgene.org Find fluorescent protein plasmids and resources Visit our Molecular Biology Reference...

  9. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    Published
    June 17, 2025, 1:15 p.m.
    ...comparison of an shRNA plasmid sequence and an miRNA plasmid sequence. (A) Plasmid sequence from pLVUHshp53... small RNA and can occur via transfection or viral delivery of a plasmid. RNAi is an umbrella term for....   Expressing shRNAs using plasmids The design of shRNA plasmids requires three important steps: ...between the red boxes. (B) Plasmid sequence from pcDNA3-miR21 (Addgene plasmid #21114). The miRNA sequence...via transient transfections and are handy for quick in vitro experiments. shRNA plasmids typically use...shRNA plasmids are being designed using RNA polymerase II promoters. The design of these plasmids is meant...with Cas13a (C2c2) Plasmids 101: The Promoter Region – Let's Go! Browse All Plasmids 101 Posts Additional...

  10. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    Published
    June 27, 2017, 1:03 p.m.
    ... create fusions with a gene of interest. These plasmids will be most useful for constructing intermolecular...ratio of acceptor to donor fusions varies with transfection efficiency and any unpaired fluorescent proteins...measure (overexpressing a gene or protein after transfection). Future of FRET The first genetically encoded...

  11. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    Published
    Dec. 31, 2019, 2:26 p.m.
    ...feature some recently deposited plasmids every quarter. The “hot plasmids” from last March were quite popular...behalf of scientists. 9. Plasmids 101: Positive and Negative Selection for Plasmid Cloning by Jennifer Tsang...changes.   1. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection by Alyssa...past year: CRISPR, optogenetics, chemogenetics, plasmids, fluorescent proteins, careers, and more. These...s the top 10 blog posts from the year. 10. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent... about mNeonGreen, mTFP1, and mWasabi and find plasmids containing these fluorescent proteins at Addgene... Read other Addgene news Find the most recent plasmid blog posts Find the most recent CRISPR blog posts...

  12. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    Published
    Jan. 10, 2023, 2:15 p.m.
    ...Addgene blog:  Plasmids 101: What is a plasmid? Plasmids 101: Origin of ReplicationPlasmids 101: PCR...process known as plasmid multimerization. Plasmid Multimerization Early studies on plasmid topology recorded...ligate the plasmid, and re-transform the ligated plasmid to recover the monomeric form of the plasmid. Have... monomer of Plasmid 12259: pMD2.G (depicted on the left) is ~5.8 kb in length. The plasmid can exist on... have been utilizing full plasmid sequencing services to verify the plasmids, including long-read sequencing... the sequencing results for their plasmids, they found the plasmids were much longer than expected, and...tandem. Such plasmids are known as multimers, and they are the result of multiple plasmid copies combining...

  13. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    Published
    Feb. 20, 2014, 4:57 p.m.
    ... smaller size of the plasmids. The Zhang lab, on the other hand, uses a 1 plasmid, single infection system... thousands of targets at once. That’s what our plasmids allow.” Wang’s library is designed to target a...target cells that have previously been stably transfected for Cas9 expression. The Yusa lab has used their...

  14. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    Published
    Dec. 14, 2020, 2:15 p.m.
    ..., cells are transfected with plasmids expressing the GPCR of interest, a Gα-Rluc8 plasmid, and the appropriate...effects.  Bryan Roth’s lab created a collection of plasmids called TRUPATH to interrogate 14 of the 16 non-visual...TRUPATH be used for? The Roth lab used the TRUPATH plasmids in a number of different experimental approaches...drugs to further understand their mechanism. These plasmids are available as a collection, facilitating screening...Addgene.org Browse the catalog of fluorescent protein plasmids Find biosensors for your experiments ...

  15. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    Published
    June 9, 2016, 2:30 p.m.
    ...for short. Addgene has already distributed this plasmid all over the world, and the question on everyone...identified NgAgo and showed that it can cleave plasmid and genomic DNA in a variety of mammalian cell ...homology-directed repair when a template is supplied. Find Plasmids from the NgAgo Publication at Addgene Cas9 and...in the cell. gDNA guides will also need to be transfected into the cells of interest. In contrast, RNA ...RNA guides must be expressed from a plasmid or in vitro transcribed, and changes in gRNA secondary structure.... Epub 2016 May 2. PubMed PMID: 27136078. Find plasmids from this publication at Addgene. 2. Swarts, Daan...24531762. PubMed Central PMCID: PMC4697943. Find plasmids from this publication at Addgene. 3. Swarts, ...

  16. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 7, 2016, 2:30 p.m.
    ... a single harvest after transfecting with the transfer and packaging plasmids. A benefit of the multiple...the passage number of a culture the lower the transfection and transduction efficiencies. Cultures should...

  17. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... CRISPR plasmids are always being added to the repository. To find all of the CRISPR plasmids available... Xiao Wang Lab has recently deposited a set of plasmids that helps identify base edited cell populations...populations without end-point sequencing assays. These plasmids use “Cas9-mediated adenosine transient reporter...and enrichment use of XMAS-TREE. a) Cells are transfected with three different vectors and flow cytometry...available from Addgene, head over to our CRISPR Plasmids and Resources page. Here are some highlights from the...

  18. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    Published
    April 5, 2022, 1:20 p.m.
    ...important collection.  Creating the Plasmids To begin constructing the plasmids for expression of mAbs, the Trimmer... This method was chosen because cells transfected with a plasmid to express the target antigen could be...expression plasmids were DNA sequenced to determine the VH and VL sequence. These plasmids have been deposited...scientific community with the plasmid sequences available from each plasmid webpage. Importantly, the methods...Resources on the Addgene blog:  Plasmid-Based Recombinant Antibodies  Plasmids 101: Secondary Nanobody Toolbox...NABOR are recombinant antibodies expressed from plasmids that have been deposited with Addgene. Here, we... first antibodies distributed by Addgene.  The plasmids used to generate these antibodies were based off...

  19. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    Published
    Sept. 14, 2017, 12:40 p.m.
    ...Harrington et al tested GeoCas9’s editing skills by transfecting HEK293 cells with GeoCas9 or SpCas9 RNP’s and...you want to give GeoCas9 a try, as always, the plasmids are available from Addgene. Happy CRISPRing! ...Background with Addgene’s CRISPR Guide Page Find CRISPR plasmids at Addgene ...

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

    Type
    Blog Post
    Published
    Dec. 10, 2013, 3:13 p.m.
    ...to sort the cells after the plasmids have been introduced by transfection or electroporation. All you ...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...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...

  21. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ...more cells = more plasmid. Plasmids can vary in copy number, the number of plasmid copies supported within...cloning, transfection, or long-term storage. But what about those pesky situations where your plasmid yield...sub-optimal plasmid yields from your prep, try our tips below! How to optimize your plasmid prep We'll ...genotype, as these mutations improve plasmid stability and yield. If a plasmid is prone to recombination Use ... a strain optimized for plasmids with repeat elements, such as viral plasmids with inverted terminal repeats...co-purify with your plasmid DNA and lead to degradation of your prep. Many plasmid prep kits recommend...levels of protein, not plasmid DNA. If you need large amounts of DNA from a plasmid in a protein expression...

  22. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    Published
    Jan. 8, 2019, 1:54 p.m.
    ...and an envelope expressing plasmid, such as our second most requested plasmid of all time, pMD2.G. Find...wide range of applications. This plasmid was the top requested plasmid of 2017. Find pMD2.G. pSpCas9(BB... top requested plasmid from 2016. You may notice the “V2.0” at the end of this plasmid which indicates...prior version of this plasmid. Indeed the first version is the 15th most popular plasmid of all time, but ...envelope expressing plasmid pMD2.G. Find pMDLg/pRPE. scramble shRNA - This plasmid from David Sabatini....1 protocol. Pro-tip: this plasmid grows more slowly than standard plasmids. Find pLKO.1 - TRC cloning...15 years of plasmid sharing has certainly taken us on many adventures...From moving office locations ...

  23. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    Published
    March 19, 2019, 1:31 p.m.
    ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... recent plasmid deposit please sign up here.          Listen to this episode of Hot Plasmids! Optogenetic...Grefen and Blatt, 2012). The 2in1 Plasmid Toolkit consists of 4 plasmid toolkits generated with 4 different...deposited the resulting optogenetic plasmid tools which includes plasmids encoding a fusion between cPYP and...targeting and editing experiments. Find the CASANOVA plasmids here! Bubeck F et al., Nature Methods 2018. PubMed...as mice, C.elegans, and Zebrafish. Find Archon1 plasmids at Addgene! Piatkevich KD et al, Nat Chem Biol...

  24. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...toolbox. Natural plasmids, also called environmental plasmids, are all around us! A plasmid is a small piece...colicin-producing plasmids have been referred to as “Col plasmids” or “Col factors.” Virulence plasmids: microbial...Old Solution but a New Problem Plasmids 101: What is a Plasmid? Plasmids 101: Transformation, Transduction...F. (2023). Plasmid classifications. Plasmid, 126, 102684. https://doi.org/10.1016/j.plasmid.2023.102684...., 2001) Beneficial traits of natural plasmids Natural plasmids often convey a variety of functions that...functions of natural plasmids. Created with BioRender.com. Resistance (R) plasmids: microbial defense...example of a plasmid benefiting its host. For example, cells with a penicillin resistance plasmid can grow...

  25. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    Published
    June 6, 2019, 12:41 p.m.
    ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... recent plasmid deposit please sign up here.      Listen to this episode of the Hot Plasmids podcast ... (ARG) plasmids to better monitor gene expression in vivo using ultrasound. These ARG plasmids contain...responses in C. elegans. Find the split BoTN/B plasmids at Addgene. Liu Q et al., Neuron. 2019. PubMed...potentially problematic for imaging. Find the ARG plasmids at Addgene. For more details on acoustic reporters...s collection consists of a library of reporter plasmids, each containing two fluorescent proteins and ...

  26. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    Published
    March 3, 2020, 2:15 p.m.
    ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...1: The base editor is split into two plasmids and co-transfected into cells for reconstitution. Image ...need. If you'd ever like to write about a recent plasmid deposit please sign up here.        AAV delivery...overcome this overlap, Yiyang Gong’s lab created two plasmids containing a red-shifted Ace-mScarlet, a fusion...channelrhodopsins. Find the Ace-mScarlet voltage indicator plasmids at Addgene! Beck et al., Sci Report 2019. PubMed...fragments. This system has been used to construct plasmids for use in model organisms such as E. coli and...

  27. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

    Type
    Blog Post
    Published
    March 24, 2016, 2:30 p.m.
    ...validated editing reagents can be found on our plasmid pages. We encourage any labs performing genome ... that are an order of magnitude greater than transfection approaches. Moreover, the cost of reagents is...CRISPR Background with Our CRISPR Guide Find CRISPR Plasmids for Your Research Check out Our List of gRNA Design...

  28. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...Neurosci 2016. PubMed PMID: 26950006 Plasmid ID Plasmid Name Plasmid Type 74309 pcDNA3.0-TRPV4-p2A-ferritin-p2A-mCherry...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These brief articles...features and applications of a partiular plasmid or set of plasmids. We hope that these articles make it ... you to find and use the plasmids you need. You can find all the hot plasmids from 2016 below. With over...over 45,000 plasmids, we can't write posts for every great plasmid that comes into the repository, but...write about your plasmids in a future blog post. No time to read? Listen to our hot plasmids segment on the...gRNA expressing cells were then transfected with a Cas9-expressing plasmid to generate a population of mutant...

  29. Working with Nuclear Receptors

    Type
    Blog Post
    Published
    July 15, 2014, 4:03 p.m.
    ...problem of sharing plasmids, providing researchers with a one-stop shop for plasmids and vectors, and making...development of a vast number expression and reporter plasmids and the free exchange of these vectors between...laboratories. The development of the transient transfection platform for assaying transcriptional activity...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 ...2005, tr22. Keep on reading! Check out Addgene's Plasmids 101 blog series. ...

  30. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    Published
    June 22, 2023, 1:15 p.m.
    ...transfer plasmid), the envelope (Env) gene on a separate plasmid (envelope plasmid), and the polymerase ... yet a third plasmid (packaging plasmid). Viral production then proceeds by transfection of all three ...different plasmids. For a retroviral system this looks like the genetic cargo in one plasmid (the transfer...three plasmids into a producer cell line, such as HEK-293T, production of viral particles, and transduction... and Pol genes are not encoded in the transfer plasmid, the component that physically integrates into ... retroviral packaging components onto distinct plasmids for safety.    Viral cargo-specific safety...services Addgene Biosafety Guide Addgene Viral Plasmids Which Type of Virus Should I Use? Resources...

  31. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    Published
    June 1, 2017, 2:30 p.m.
    ...dish of mammalian cells is transiently transfected with a plasmid containing a fluorescent protein (fusion...dish. This is caused by the stochastic nature of plasmid uptake by cells. To correct for this variation,...Goedhart et al, 2012, Bindels et al, 2017). The plasmid for co-expressing mTurquoise2 with mVenus as a ...Find Empty Backbones with Fluorescent Tags Find Plasmids for Subcellular Localization Browse the entire...

  32. RaPID Detection of RNA-protein Interactions

    Type
    Blog Post
    Published
    Aug. 16, 2018, 12:40 p.m.
    ...Split-BioID plasmids Find CRISPR plasmids for labeling genomic regions Find CRISPR plasmids for purifying... include both scrambled RNA motifs and cells transfected with BirA* protein alone (i.e. no RNA bait). ...question that RaPID could help answer? The RaPID plasmids are available from Addgene! References Ramanathan...PubMed Central PMCID: PMC5886736. Find the RaPID Plasmids Here! McHugh, C.A., Russell, P., & Guttman, M...

  33. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    Published
    Nov. 6, 2018, 1:12 p.m.
    ... [6], plasmid electroporation (episomal) [7, 8] and mRNA transfection [9]. Many of the plasmids used for...Neurons [32, 33, 34] You can find a variety of plasmids for differentiation here. Transdifferentiation...Weintraub, and A.B. Lassar, Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell,...Pages Find Viral Vectors for Your Research Find Plasmids from the Yamanaka Lab ...

  34. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  ...all applications. Addgene’s plasmid collection includes over 10,000 plasmids with a FLAG tag (!), but did...electroporation or poor cellular entry with other transfection methods. The Junwei Shi Lab addressed this problem... al. 2023 with permission.   Find Cas-PAGE plasmids here!   Zhang, Z., et al. (2023). Efficient engineering...inspired and informed this section.  Find mStayGold plasmids at Addgene here!   Ando, R., et al. (2023). StayGold...under CC-BY license.   Find the AAV.CAP-Mac plasmid here!   Chuapoco, M.R., Flytzanis, N.C., Goeden...(2023) under CC-BY license.   Find AAV1-X1 plasmid here! Chen, X., et al. (2023). Functional gene ...

  35. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    Published
    Oct. 12, 2017, 2:26 p.m.
    ...as a universal donor plasmid. The CRISPaint toolkit includes several donor plasmids for protein tagging...The Jorgensen Lab recently developed a modular plasmid assembly toolkit called SapTrap to improve CRISPR...repair template. SapTrap includes a prebuilt donor plasmid library containing several types of fluorescent...tag and homology arms). Digestion of the donor plasmids with SapI allows the tag, selectable marker, and...to be released (Fig. 1, Step 2-3). Since donor plasmids of the same type will produce the same unique ...different combinations of connector and tag donor plasmids from the library can be used to generate functionally... of the targeting vector and a Cas9 expression plasmid will insert the desired genetic tag and marker ...

  36. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...structures within cells. HeLa cells were transfected with plasmids encoding FPs tagged with the plasma membrane...They generated plasmids tagged with EB3 to mark the growing tips of microtubuli and plasmids tagged with ...and dCas9-VPR targeting plasmids can be found at Addgene. Check out these plasmids for piggyBac gRNA delivery...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head... Find the plasmids at Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting...

  37. AAV Q&A with Tim Miles

    Type
    Blog Post
    Published
    May 17, 2022, 1:15 p.m.
    ...Animal Models Q9: Is there any difference in transfection efficiency when delivering these viruses via...user Resources on Addgene.org Find Adenovirus Plasmids Adenovirus Guide Page Subscribe to Viral Vectors...

  38. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    Published
    Sept. 9, 2014, 8:20 p.m.
    ...multitude of techniques including co-transfection of two or more plasmids, the use of multiple or bidirectional...with a plasmid that already has the multicistronic element and reporter cloned in. In these plasmids you ...interest. Some of the plasmids listed in the table above (and their related plasmids) are designed to have... a one-stop reference guide for plasmids. We've combined our Plasmids 101 blog posts and additional resources...desktop. The Plasmids 101 eBook is designed to educate all levels of scientists and plasmid lovers and ... an introduction to plasmids, allowing you to spend less time researching plasmid basic features and spend...May 11.) PubMed.   Read more Plasmids 101 posts or browse Addgene's Plasmid Guide for more molecular biology...

  39. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...adenoviral stock (rAdV-S). HEK293 cells are transfected with the AdV plasmid construct from step #1 and allowed...down into 5 key steps: Construction of the rAdV plasmid (~1 week): The AdEasyTM system is the most popular...adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the transgene of interest...how to generate the final recombinant adenoviral plasmid construct, please see Addgene’s Adenoviral Guide...2.*Pro-Tip*: once the correct recombinant pAdV plasmid is identified, re-transform into a bacterial strain...recombination for amplification. The final purified plasmid stock should be analyzed by endonuclease restriction...transductions Resources on Addgene.org Find Adenovirus Plasmids Adenovirus Guide Page  ...

  40. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ...to find Gateway plasmids for your organisms of interest: Find Gateway cloning plasmids here! References...antibiotic resistance markers (indicated here by plasmid color), allowing you to easily select for the expression...desired, recombined construct. Read our recent Plasmids 101 Post on CcdB for more information. How to ...Method A: recombination of an attB-PCR product or plasmid with an attP donor vector. In this case, we would... to use the Snapgene program to design Gateway plasmids.    Figure 3: Method A to create an entry...restriction digest! Then, you can transform or transfect the cells that you want to use for your experiments...entry clones can be used with a large variety of plasmids. Speed The Gateway system enables the generation...
Showing: 41 - 80 of 97 results