...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...

...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 ...

...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...

...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...

...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 ...

...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 ...

...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...

...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...

... 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...

...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 ...

...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...edits in protoplasts or plants of rice and wheat. Read the paper in Genome Biology Find plasmids containing...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...paper in Nature Communications Find the Cas12b plasmids  Plant base editing with a Cas9-adenosine deaminase...Neuroscience Find the AAV-based SaCas9 genome editing plasmids String assembly gRNA cloning (STAgR) String ...the paper in PLoS One Find the STAgR cloning plasmids Mobile-CRISPRi allows CRISPRi experiments in...Gross’s lab recently deposited “Mobile-CRISPRi” plasmids, a set of modular CRISPRi tools that can be used...

...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids Topics: CRISPR, Other CRISPR... where we highlight a few of the newest CRISPR plasmids available at Addgene. If you want to read more...will cover: Miniature CRISPR-Cas systems SEND plasmids Prime editing improvements Miniature CRISPR-Cas...Read the paper in Molecular Cell Find the CasMINI plasmids Efficient CRISPR editing with a hypercompact ...paper in Nature Biotechnology Find the Cas12f-GE plasmids Programmed genome editing by a miniature CRISPR-Cas12f...cells using various delivery methods, including plasmid, ribonucleoprotein, and AAV.  Read the paper in... in Nature Chemical Biology Find the AsCas12f1 plasmids   Selective endogenous encapsidation for cellular...

... As reviewed in our Plasmids 101 series, proteins are overexpressed from a plasmid construct, most often...multiplexing. pCri plasmids are divided into two sets, each of which includes plasmids with different varieties...on the Addgene Blog Plasmids 101: E. coli strains for protein expression Plasmids 101: Protein tags ...varieties of tags, both N- and C-terminal. Each set of plasmids uses the same restriction enzymes for cloning ...for one protein quickly and easily. These tagged plasmids also include cleavage sites should one need to...other tags used in the pCri system, please see Plasmids 101: Protein Tags. Beyond E. coli: purifying ...subtilis and Pichia pastoris pCri also includes plasmids designed for protein expression in systems other...

... by 10% plasma, but GeoCas9 remained active in up to 30% plasma. Figure 2: Human plasma longevity...activity in HEK293’s. GeoCas9 stability in human plasma The last trick GeoCas9 has up its sleeve is its increased...diluted human plasma. GeoCas9 remains active after being incubated in up to 30% human plasma. The discovery...incubated SpCas9 and GeoCas9 RNPs in diluted human plasma for 8 hours at 37°C. Substrate was then added and... SpCas9 or GeoCas9 RNPs were incubated in human plasma diluted with reaction buffer for 8 hours at either...excited about? Thermostability? Longevity in human plasma? Something we haven’t thought of yet? Let us know...you want to give GeoCas9 a try, as always, the plasmids are available from Addgene. Happy CRISPRing! ...

...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...called a bacmid. Like AAV and lentiviral packaging plasmids, bacmids are readily available, so the amount ...insect and mammalian systems. Baculovirus transfer plasmids have a much larger packaging capacity — up to ...can produce AAV and lentivirus if needed # of plasmids Three or four Two Packaging limit ~4.7 kb (AAV...

...Here are some of the AAV we have released in the last few months. You can also browse our entire AAV inventory...constructs from Christopher Harvey’s lab. These plasmids localize opsins to the soma to avoid off target...neighboring neurons. We have also packaged hChR2(H134R) plasmids from Karl Deisseroth’s lab, which includes pAAV-hSyn...H134R)-EYFP that is both Cre and Flp dependent. Plasmid Serotype Name 124603 AAV9 pAAV-EF1a-DIO-ChrimsonR-mRuby2... has now expanded to offer many of our control plasmids in new serotypes. See our Serotype Testing AAV...AAV inventory for the entire collection! Plasmid Serotype Name 51502 AAV5 pCAG-FLEX-EGFP-WPRE 114471... jGCaMP7 from Janelia, including three jGCaMP7 plasmids now packaged in AAVrg. jGCaMP7s, the variant with...

...features was our collection of popular plasmid sequences. These plasmids contain features such as antibiotic... our SnapGene powered plasmid maps Learn how we use SnapGene during the plasmid quality control process...out full plasmid sequencing at Addgene Additional Resources on Addgene.org Example plasmid analysis ...design was done painstakingly by hand. Records of plasmid construction were often incomplete or nonexistent...popular aspects - its ability to annotate a raw plasmid sequence and display frequently used genes and ...when the DNA sequence has changed. Identifying plasmid control elements Coding sequence features are relatively... boundaries for control elements in commercial plasmids were inconsistent and sometimes clearly wrong....

...genome is lethal. If this DSB occurs on a plasmid, the plasmid will be eliminated from the bacterium, which...sequence-specific, antimicrobial, plasmid-based, CRISPR systems (Find the Citorik Plasmids here). In both cases, ...genes in E. coli using plasmid and phagemid delivery systems. Phagemids are plasmids containing a phage origin...sequence results in cell death. Plasmid targeting may result in plasmid loss and antibiotic resensitization...this resistance gene being encoded by a high-copy plasmid. In the case of the genomically encoded quinolone...RNAs (crRNAs) targeting either genomic (blue) or plasmid-based sequences (red). Treating a mixed population...specifically targets bacteria with the given genomic or plasmid sequence from a mixed population without affecting...

... after ordering a plasmid you didn't have to grow the bacteria and prep the plasmid before you begin your...with 210 plasmids from the collection. You can find the full list here. Most of these plasmids are empty...cloning experiment? What if after receiving the plasmid from Addgene you could directly start your digest...you can focus on what you want to do with your plasmid without spending time and resources in preparative...research and discovery.” Here are the details. Plasmids offered as cloning grade DNA As we begin testing...molecular cloning. Plus, we decided to include 10 plasmids that can be directly used in the research on the... the SARS-CoV-2. Nine of the overall plasmid selection will be open to industry scientists. This collection...

... that made it easy to share plasmids, both for people depositing plasmids and for people requesting them... Collections Lentiviral Plasmids Tetracyline Inducible Plasmids Empty Backbones Bacterial Expression...Expression Systems AAV Plasmids Blogs Origin of Replication What is a Plasmid? How to Design Your... Verify Your Plasmid Using a Restriction Digest Analysis Videos What is a Plasmid? How to Design...video, however, is “What is a Plasmid?”, An animated explainer of a plasmid and its parts that helps the...them. Prior to this, many plasmids were available, but they often weren’t easily accessible, due to a ...barriers, allowing people to access a wide catalog of plasmids from all over the world. This idea, that available...