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We narrowed to 416 results for: plasmids 101

Showing: 241 - 260 of 416 results

  1. DNA Purification Without a Kit

    Type
    Blog Post
    ... Read our Plasmids 101 blog series Find other plasmid protocols and tips Browse our plasmid cloning blog...alkaline lysis plasmid miniprep Starting material: 2 mL bacterial culture Product: plasmid DNA This kit-...kit-free plasmid miniprep protocol from Addgene follows a similar workflow as a column-based plasmid extraction... or agarose gel slices Product: plasmids or DNA fragments The plasmid purification and DNA gel extraction...depending on the type of DNA you’re purifying (plasmid, genomic, or DNA fragments from agarose).    In...causes the proteins and genomic DNA to precipitate. Plasmid DNA is free in solution. Proteins and genomic DNA... removed by centrifugation. At this point, the plasmid DNA sample can be treated with RNase to remove ...

  2. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...Fusions Plasmids 101: Secondary Nanobody Toolbox Resources on Addgene.org View the Antibody plasmid collection...The Seeger lab found a way around this by using plasmids to generate sybodies, synthetic nanobodies against...nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target epitopes that...form of mRNA for academic research. The template plasmids for generating your own libraries are also available...available in the Sybody Generation Toolbox plasmid kit: SB_concave, SB_loop, SB_convex. These scaffolds...efficiently subcloned into the phagemid pDX_init plasmid, using the fragment exchange (FX) cloning technique... subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for the production of tag-free proteins for further...

  3. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...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...

  4. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...resources on the Addgene blog Read all of our Plasmids 101 blog posts Read our blog post on 5 factors to...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...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...

  5. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    ...Fluorescent Protein 101 blog series Learn more about FRET Find protocols and tips for making plasmid constructs...at the cells on the plate. For example, if your plasmid contains a gene encoding a blue chromoprotein, ...colonies to determine which cells have taken up the plasmid. Of course, other follow up methods like sequencing... chromoprotein sequences into E. coli BioBrick plasmids (meffRed, eforRed, asPink, spisPink, scOrange,...chromoproteins. In some cases, cells containing high-copy plasmid-borne chromoproteins lost color overnight in liquid...problem. There is no single “ideal” chromoprotein plasmid that combines the most desirable features: intense...Chromoproteins for allelic exchange Figure 3: pTOX plasmids expressing amilCP and tsPurple genes. Image: Lazarus...

  6. New and Upcoming Viral Vectors - September 2019

    Type
    Blog Post
    ...is now complete! pAAV-CAG-GFP (plasmid 37825) and pAAV-hSyn-EGFP (plasmid 50465) are now available as 20...tips for preparing AAVs Download our viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's...

  8. Viral Production at Addgene

    Type
    Blog Post
    ...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral.... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ...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...undergoes PCR with primers targeting the transfer plasmid used in the transfection. PCR products are analyzed...microbial contamination. Purity When possible, all plasmids used for viral production are propagated in the... system and PCR-based confirmation of transfer plasmids used in Addgene’s viral production. Barcoding ...

  9. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...Inside Addgene Posts! Learn all about Plasmids in our Plasmids 101 series! ...bacteria with the plasmid can grow. As the bacteria divide, they replicate and retain the plasmid. This allows...Every plasmid sample enters Addgene the same way. A package is delivered by a mail courier and then the...about Developing Lab Management Software    The plasmid's journey begins First, each package is opened and... to hundreds of samples, with an average of 10 plasmids per deposit. Each week an average of 16 deposits...allow us to store multiple types of tube for each plasmid we have at Addgene. Each sample at Addgene has ...strains that have been treated to easily take in plasmids (transformation). Transformed bacteria are then...

  10. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...mutagenesis of Plasmids. A) Generation of mutagenized plasmids. PCR primers (green) amplify the plasmid template... Cloning Techniques Learn More about Plasmids in Our Plasmids 101 Series Use CRISPR to Generate Specific...with the nuclease-resistant nicked plasmid (the PCR product). Plasmids are isolated from the resulting colonies...the final circular plasmid product pictured here. B) Screening of recovered plasmids for the mutation of...Smaller plasmids (~3 kb) are generally more efficiently amplified than larger constructs, but plasmids as ...DpnI) to eliminate the parent plasmid from the PCR products, the plasmid template should be isolated from...have taken up the mutagenized plasmid. Note that any residual parental plasmid (usually from incomplete DpnI...

  11. OCaMP: A New Calcium Indicator for Neural Imaging

    Type
    Blog Post
    ...doi.org/10.1101/2025.07.28.667269. Additional Resources on the Addgene Blog Viral Vectors 101: Calcium...imaging, there's a version that fits your setup: Plasmid Backbone Promoter pRSET OCaMP pRSET ...button_text" is_json="true" %}{% raw %}"Find OCaMP plasmids available at Addgene!"{% endraw %}{% end_module_attribute...on addgene.org Fluorescent Proteins: Biosensor Plasmids In-Stock AAV Vectors: Biosensors AAV Packaged ...

  12. Typing CRISPR Systems

    Type
    Blog Post
    ...https://doi.org/10.1016/j.ggedit.2022.100013 Additional resources on the blog CRISPR 101: Cas9 vs. The ...recently that researchers began building optimal plasmids to harness the power of Class 1 systems. Class...Lastly, type IV systems are most often found on plasmids. The many unique features of IV-A and IV-B have...led to the hypotheses that they are involved in plasmid competition or can hijack machinery from other ...highly diverse and involved in competition between plasmids. Nucleic Acids Research, 48(4), 2000–2012. https...The Other Cas(s) CRISPR 101: RNA Editing with Cas13 Build your CRISPR vocabulary Additional resources on... on Addgene.org CRISPR guide CRISPR plasmids CRISPR references ...

  13. MXS Chaining

    Type
    Blog Post
    ...Gibson Assembly Learn About Other Plasmid Cloning Techniques Plasmids 101: Green Fluorescent Protein Resources...Fluorescent Protein Collection Find Plasmid Kits at Addgene Find Plasmid Cloning Protocols References 1....combination of different genetic sequences into plasmid DNA. In high throughput cloning techniques, although...MXS chaining MXS-chaining was designed to create plasmids for fluorescence imaging or flow cytometry applications...modules (Figure 2) are contained in individual plasmids and are each flanked by the same multiple cloning...throughput cloning is to facilitate the construction of plasmids with many components. The creators of the MXS-... plasmid cloning...

  14. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...sequencing on select regions of all distributed plasmids as part of quality control, but does not perform...unlike an RNA guide, it cannot be produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo...with 5’-P-ssDNA guides and an NgAgo expression plasmid to edit a gene in vivo. Try NgAgo for Yourself ...complexes or with guide RNA- and Cas9-expression plasmids. Cas9 guide RNA delivery does however rely on ...(1), NgAgo has received much attention and the plasmid has been requested by labs around the globe nearly...happy genome editing! Check Out Other Argonaute Plasmids from the Doudna Lab Many thanks to our Guest .... When not snipping DNA, he tries random things (101 lessons on Spanish, C++, guitar, photography) and...

  15. Working 9-5: an Addgene Co-op

    Type
    Blog Post
    ... the submitted sequences were a concern to the plasmid function. The steps of the training process gave...parts of Addgene. This was done through the “Addgene 101” series, where each team gives a presentation on ...

  16. DAP arrays for multiplex gene editing

    Type
    Blog Post
    ...tRNA-gRNA arrays, which is scalable and efficient in plasmid and viral vectors. In applications when repetitive...manual and online design examples of using Addgene plasmids hCtRNA_FT (#186715) and hCtRNA_VT (#186716).   ...Resources More resources on the Addgene blog CRISPR 101: Multiplex expression of gRNAs Four Base Editing ...

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

    Type
    Blog Post
    ...Drosophila Read our Plasmids 101 blog series Resources on Addgene.org Find plasmids for your research ... 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...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...

  18. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...infiltrate cells. Options for CRISPR delivery include plasmid transfection, RNP electroporation, and viral transduction...types they tested. Interested in using Nanoblades? Plasmids are available from Addgene. Genome editing primary...to DNA transfection of gRNA and Cas9 expression plasmids. These results are in line with previous research...delivery methods for mammalian cells Browse our CRISPR 101 blog post Resources on Addgene.org Check out our...our comprehensive CRISPR guide Find CRISPR plasmids and resources Learn about viral production ...

  19. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ... 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...

  20. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...transformation Read our Plasmids 101 blog posts Resources on Addgene.org Browse our Plant Plasmids and Resources...page Find synthetic biology plant plasmids Find CRISPR plant plasmids ...its function. In nature Agrobacterium contains a plasmid with a T-DNA (transfer-DNA) region. During infection...localization of your protein of interest. Find all plasmids from the Lee-Parsons lab. Test different leaf...used the disarmed Agrobacterium tumefaciens strain GV3101(pMP90), which works well for us. Use a well working...
Showing: 241 - 260 of 416 results