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We narrowed to 1,038 results for: LAS

Showing: 221 - 240 of 1038 results

  1. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ...transfer plasmid), the envelope (Env) gene on a separate plasmid (envelope plasmid), and the polymerase ...different plasmids. For a retroviral system this looks like the genetic cargo in one plasmid (the transfer...Gag structural protein on yet a third plasmid (packaging plasmid). Viral production then proceeds by transfection... transfection of all three plasmids into a producer cell line, such as HEK-293T, production of viral particles... 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.... (AAVS1)). For these reasons, AAV is usually classified as a BSL-1 safety level. Adenovirus is similar...

  2. Simplify Cloning with in vivo Assembly

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    Blog Post
    ...blog posts on plasmid cloning Resources on Addgene.org Read our protocols for plasmid cloning Find our... from the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential part of any molecular biology... techniques involve the assembly of a circular plasmid in vitro, before transforming it into E. coli for... propagation. However, while not widely known, plasmid assembly can be achieved in vivo using a bacterial...cloning tool was first reported over 25 years ago (Douglas and Howard, 1991, Bubeck et al., 1993). But why...can be encoded in the primer sequences. Multiple plasmids can be included in a single PCR, for example for...linear fragments are transformed into E. coli and plasmid assembly (re-circularization) occurs in vivo. We...

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

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    Blog Post
    ...encoded on separate plasmids and are co-transfected into cells alongside a plasmid encoding the minigenome...four viral genes in the same plasmid (as opposed to four separate plasmids), there are a few reasons arguing...separate (and thus smaller) plasmids. Finally, having four separate plasmids allows researchers to partially... 2 μg minigenome plasmid per well (in a 6 well plate format). However, since plasmid quality and transfection...Minigenome System). From a more technical standpoint, a plasmid with all four genes and necessary control elements...relatively large. Cloning the viral genes in this plasmid would be more difficult compared to if each gene...Technical tips It is important that all of these plasmids (those containing NP, VP35, VP30, L, and the minigenome...

  4. CRISPR 101: Any Base Transversion Editors

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    Blog Post
    ... N-methylpurine DNA glycosylase (MPG), also known as alkyladenine DNA glycosylase (AAG). When adenine ... MPG / AAG N-methylpurine DNA glycosylase / alkyladenine DNA glycosylase  Removes hypoxanthine from DNA...In our last post, we talked about the first base transversion editors: CGBEs, or C → G Base Editors. ...using either fused or endogenous Uracil DNA N-glycosylase (UNG) enzymes. Various other proteins included... (Y = T or C) base editor, which they called glycosylase-based guanine base editor (gGBE) (Tong, Liu, ...Editor Converts C → G by fusing a uracil DNA glycosylase to a CBE; the first type of base transition editor...apyrimidinic (AP) site. UNG / UDG Uracil DNA N-Glycosylase Removes uracil from DNA, creating an abasic site...

  5. Addgene: The First Twenty Years

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    Blog Post
    ...about the amazing years in front of us. So in our last celebratory moment, we wanted to walk through some...on Twitter. In the beginning Let’s take a quick flash back to 2004. “Shrek 2” was the most popular movie...frustrated by the difficulty of accessing published plasmids from other scientists. She, Kenneth Fan, and Benjie...nonprofit that would facilitate the sharing of plasmids between scientists. The service would allow people...Addgene before we were a “labhold” name. Our first plasmid request came on September 8, 2004, for pcDNA-p160MBP... our first major repository milestone — 10,000 plasmids deposited and available — and nearly seven before...possible for scientists to both deposit and request plasmids, was a major focus of our first decade. We did...

  6. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ...summer or just a bright new plasmid. One area with lots of bright new plasmids is our Fluorescent Biosensors... for AAV packaging.    Find iGECInano plasmids here!   Matlashov ME, et al. Design and Initial Characterization... latest additions to Addgene’s catalog is a new class of sensors for neuropeptides and neurotransmitters...SF-iGluSnFR. Image from Aggarwal 2022.   Find iGluSnFR3 plasmids here! Find viral prep here: pAAV.hSyn.FLEX.iGluSnFR3.... 2022, under CC-BY-NC license.   Find JEDI-2P plasmids here! Find viral preps here: pAAV-EF1a-DIO-JEDI...signaling Receptor tyrosine kinases (RTKs) are a major class of signaling receptors, but the options for measuring...reporter, they observe fluorescence localizes to the plasma membrane only when the RTK is activated and the...

  7. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...given protein? Classical methods for studying protein phosphorylation A number of classic methods exist...expression plasmid encoding the ORF to be expressed Finally, the researcher needs a plasmid encoding their...#68306 pCRT7 NT TOPO E17TAG GFP plasmid - Addgene #68295 SupD plasmid – Addgene # 68307 For further technical...Find Synthetic Biology Plasmids Find Bacterial Expression Systems Find Plasmids for Protein Purification... methanogenic archea: these organisms lacked a classic cysteinyl tRNA synthetase, despite abundant incorporation...strain for protein production, and an expression plasmid encoding the open reading frame (ORF) of interest...translation of pSer-incorporated proteins on a single plasmid. This includes 4 copies of a tRNA that specifically...

  8. PEI Calculator for Planning AAV Packaging Transfections

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    Blog Post
    ...names of the plasmids you plan to use Enter the base pair size of those plasmids Enter plasmid concentration...ratios below: 1 Transfer Plasmid : 1 pHelper: 1 RepCap OR 3 Transfer Plasmid : 5 pHelper: 2 RepCap Select...of each plasmid needed per dish or stack. Calculations are based on using 28 μg of total plasmid (Transfer...per 15 cm dish or 2500 μg of plasmids per CF5 stack. Total ug of each plasmid needed for the whole experiment...packaging typically requires transfecting three plasmids at specific molar ratios. To get these ratios ...transfection calculator for AAV packaging Select the plasmid molar ratio you would like to use for your AAV ...experiment Total volume (μL) of plasmid needed for the whole experiment And here’s the equation the calculator...

  9. New and Upcoming Viral Vectors - June 2019

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    Blog Post
    ...Chemogenetics Plasmid Serotype Name 119472  AAV5  AV SYN PSAM4 GlyR IRES EGFP   Biosensors Plasmid Serotype...Here are some of the AAV we have released in the last few months. You can also browse our entire AAV inventory...inventory of serotype testing AAV New control AAVs Plasmid Serotype Name 105530  AAV1 pAAV.CMV.PI.EGFP.WPRE.bGH...biosensor AAV inventory New glutamate sensors Plasmid Serotype Name 106176  AAV1  pAAV.hSynapsin.SF-iGluSnFR.S72A...pAAV.hSynapsin.SF-iGluSnFR.S72A New calcium sensors Plasmid Serotype Name 104496  AAV1 pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE...pAAV-hSynapsin1-FLEx-axon-GCaMP6s New voltage sensors Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST...of DREADDs / Chemogenetics AAVs New DREADDs Plasmid Serotype Name 75033  AAV1 pAAV CD68-hM4D(Gi)-...

  10. CRISPR/Cas9 FAQs Answered!

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    Blog Post
    ...easiest way is to co-transfect several plasmid together, with each plasmid having a targeting spacer cloned ...articles (3/12/2014). With so many options for CRISPR plasmid tools and numerous experimental design decisions...with expression, though it is possible for the plasmid to still express without the G. Because the G is...experiments for multiplexed targeting with Zhang lab plasmids? A6: For multiplexing CRISPR to target multiple...into PX330, then co-transfect or delivery the two plasmids together into your cells. If you want to use double...very big concern, you could go ahead with PX330 plasmid, clone in your target guides, test run them, and...donor template, it's usually more efficient than plasmid donor. Typical design is to have 50–80 bp of homology...

  11. New and Upcoming Viral Vectors - May 2020

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    Blog Post
    ...our control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression...AAV and lentiviral preps we have released in the last few months: Lentiviral preps for expressing SARS-CoV...protein expression The Krogan lab quickly shared plasmids expressing SARS-CoV-2 proteins at the beginning.... Now, we've turned some of the most requested plasmids into lentiviral preps to speed up COVID-19 research...Testing AAV inventory for the entire collection! Plasmid Serotype Name Depositor 27056 AAVrg pAAV-Ef1a-DIO...constructs that allow for targeted expression. Plasmid 75470 is now available in AAV1 for Flp-dependent... mCherry reporter. We now also offer AAVrg for plasmid 83898, which targets expression of ChR2 to forebrain...

  12. Quick Guide to All Things Lentivirus

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    Blog Post
    ...virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing all the structural genes...crossover event between the vector plasmid and the packaging plasmid caused the Psi sequence to be placed...expression of the transgene   Transfer plasmids The transfer plasmid is the vector you clone your gene of...usually 293T cells, with your transfer plasmid and your packaging plasmids. For this, you can use the transfection...techniques are still used and efficiently transfer plasmid DNA efficiently into dividing immortalized cells... (Gag, Pol, Tat, Rev and Env) and the transfer plasmid encoding the insert of interest as well as sequences...the packaging signal Psi). Because the packaging plasmids lacked the psi signal, the genes encoding the ...

  13. The Stingy Scientist: How the Baby Gel Box Was Born

    Type
    Blog Post
    ...science project was to make my own glass milk for purifying cut plasmid DNA from agar slices.  Molecular...me that most Addgenies make an effort to avoid plastic bags and to conserve resources where possible.  ....  At the lunch table, for instance, we have a plastic container problem…reusable containers pile up by...sink after lunch as fewer and fewer people use plastic bags.  We also try to recycle our discards according...about conserving resources in the labs. Disposable plastic materials are fairly inexpensive in this country... funded and we didn’t feel the need to reuse plasticware or think about conserving reagents and buffers...lifetime supply for about $10 US dollars worth of glass powder pruchased from a pottery supply house.  These...

  14. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...are excited by the yellow/green laser but only "bright" for the blue laser. To understand why that is, let's...brightness changes to "dim" for the same laser type. Knowing that the red laser excites at 640 nm (Table 1), while...separately detect these markers in your analysis. Lastly, there are fixable dye stains that react with free...few commonly used fluorophores in relation to the laser type. Table adapted from: https://emea.bd.com/panel-design...-fluochrome-brightness-spillover Excitation Laser Color Very Bright Bright Moderate Dim ... brightness beyond the fluorophore itself, like laser type and power, acquisition voltage, the concentration... let's look at Figure 2. The blue laser excites at a wavelength of 488 nm (Table 1), which is near a local...

  15. Fluorescent Tagging of Endogenous Genes with SapTrap

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    Blog Post
    ...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 ...

  16. Depositor Week 2020 is Happening Now. Join us!

    Type
    Blog Post
    ...While the bulk of these plasmid deposits came in this year, there were some plasmids deposited as early as... We started from 474 plasmid deposits in 2014 which has grown to ~10,000 plasmid deposits per year in ...the scientific community the latest and greatest plasmid tools! Thank you depositors! In fact, this year...quickly. There are over 2,300 COVID-19 related plasmids currently in Addgene’s repository which have been...the GSA’s blog Genes to Genomes about a TMPRSS2 plasmid from Roger Reeve’s lab as an example). Depositor... of the Addgene community. Perhaps you’ll find plasmids and new tools that will speed your science along...and trivia begins! Tuesday: Depositor features (plasmid deposits and blog posts showcasing the work of ...

  17. CRISPR 101: Ribonucleoprotein (RNP) Delivery

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    Blog Post
    ...more stable in human plasma than the standard spCas9 (Harrington et al. 2017). Lastly, as with any scientific...commonly performed by transfecting cells with a plasmid, such as PX459, which encodes Cas9 and contains...chosen for both Cas9 and gRNA expression       Plasmid DNA may be incorporated into the genome     Off-target...targets with similar efficiency as compared to plasmid-based expression of Cas9/gRNA and can be used for... as CMV or EF1A promoters found in many CRISPR plasmids) are not expressed. Because this method does ...fluorophore tagged dCas9 may require the use of plasmid or viral-mediated delivery. You’ve decided to...and purify His-tagged Cas9 from E. coli using a plasmid such as pET-28b-Cas9-His from Alex Schier’s Lab...

  18. Multiplexed Overexpression of Regulatory Factors (MORF) collection

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    Blog Post
    ...Addgene.org Plasmid Pooled Libraries Pooled library amplification Zhang Lab CRISPR Plasmids The Human TFome...MORF Collection) and an accompanying expression atlas, including over 3,500 human TF isoforms, to help...understand this regulatory network. The transcription atlas and library The transcription factor library The...determine how its presence influences cell fates. The atlas of directed differentiation To understand how each... fates. (Adapted from Joung et al.)    This atlas linked TF expression to differentiation outcomes...characterized TFs, the Zhang group were able to classify unknown, or ‘orphan’, TFs into their respective...cell types generated by the human fetal expression atlas. Computational analysis was used to infer differentiation...

  19. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

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    Blog Post
    ... 1998). Interested in plasmids that contain the groucho gene? Check out plasmids deposited by the Shivdasani... several plasmids that contain SHH, you can find them all here! The most popular SHH plasmid however was... to see for yourself. We have one plasmid in our repository, plasmid 39624, that contains cmr.   tinman...website for plasmids that contain tinman or the mammalian homolog Nkx2.5. We also have one plasmid, Addgene...Drosophila Read our Plasmids 101 blog series Resources on Addgene.org Find plasmids for your research ...the stories behind the names. We also have many plasmids in our repository that you can use to study these... Beauty (Ivics et al., 1997). We have a lot of plasmids that contain components of the SB transposon system...

  20. 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...Otherwise, check out our plasmid and DNA backgrounds. Download the blue plasmid background.     You can... the plasmids that Addgene offers by visiting our collections page. Here, you can browse plasmids by field...your favorite reagent, scientific technique, or plasmid repository at your next meeting.     If you’re...cute mascots!  Blugene has been helping us make Plasmids 101 videos and needed a break so they're hanging...coronavirus collection page, where you can find plasmid tools to accelerate your COVID-19 research.  Download...background.         If you’re predominantly a plasmid user, instead, don’t fret; we have backgrounds ...
Showing: 221 - 240 of 1038 results