We narrowed to 383 results for: plasmid dna
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TypeBlog PostPublishedJune 26, 2018, 1:33 p.m....working with the Rare Genomics Institute to provide plasmid grants to researchers working on rare diseases ...Richard Gronostajski who each are being awarded free plasmids form Addgene to accelerate their research. Read... on to learn how these researchers plan to use plasmids from Addgene to advance their research on diseases...Prof. Videira and collaborators will use Addgene plasmids to immortalize CDG patient-derived immune cell...understanding the structural determinants of protein-DNA interactions and how these interactions regulation...Nfix-negative neural stem cells. The lab will use plasmids from Addgene to construct NFIX expression vectors...
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An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency
TypeBlog PostPublishedDec. 17, 2020, 1:30 p.m....CRISPR/Cas9 ribonucleoprotein complex with linear DNA as repair templates which reduces time to generate...repair templates with homology arms of 35-80 bp, and DNA double-strand breaks up to 54 bp from the desired...in knock-in efficiency, presumably by channeling DNA breaks into the homologous recombination repair pathway.... elegans. These modular toolkits build single plasmid targeting vectors that encode the gRNA transcript... C. elegans, visit Addgene’s C. elegans CRISPR plasmids page. Future perspectives It is interesting to...Single-Stranded and Long, Partially Single-Stranded DNA Donors in Caenorhabditis elegans. Genetics 210:781...5′ Modifications Improve Potency and Efficacy of DNA Donors for Precision Genome Editing. BioRxiv. https... -
Plasmids 101: Biotinylation
TypeBlog PostPublishedNov. 15, 2018, 1:50 p.m....biotinylating proteins with BirA. You can find plasmids that contain both AviTag and BirA at Addgene! ...labeling Biotin can be added to either terminus of a DNA sequence during a Polymerase Chain Reaction (PCR)...Photobiotin can be used to non-selectively label DNA and RNA. Photobiotin acetate consists of a biotin...electrophoretic mobility shift assay to examine specific dsDNA, ssDNA or RNA–protein interactions.” Nucleic Acids ... -
Visualizing Genomic Loci with CRISPR-Sirius
TypeBlog PostPublishedApril 18, 2019, 12:53 p.m....Addgene.org Browse our CRISPR plasmids for visualization Find plasmids for your research Read our CRISPR...protein (dCas9) is well known for its ability to bind DNA targets without changing them. Thus, it has been ...cells. Based upon this, they chose an intergenic DNA region (IDR3) as a reference loci to measure inter-locus..., give CRISPR-Sirius a try. Find CRISPR-Sirius plasmids at Addgene. References Ma, Hanhui et al. “Multicolor... -
Tips for CRISPR Gene Editing in Mice
TypeBlog PostPublishedJune 28, 2016, 10:59 a.m.... target plasmid and pX330-sgRNA plasmid (Mashiko et al. 2013). The pCAG-EGxxFP target plasmid contains...pCAG-EGXXFP plasmid along with the individual gRNAs in their PX330 plasmids (or similar plasmids for gRNA... (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences ...degradation that can set you back days. Working with plasmid DNA is much easier and doesn’t require the extra ... system floating around, and many pre-designed plasmids containing these variants ready for purchase. ... or you can order them yourself through whatever DNA supply company you currently use. You can also find...multi-cloning site (MCS) between EGFP fragments. The pX330 plasmid contains humanized Cas9 expression cassette and... -
Hot Plasmids: Fall 2024
TypeBlog PostPublishedNov. 21, 2024, 1:30 p.m....of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. ...indels after DNA repair. B) M. abscessus is first transformed with Cas9 on an integrative plasmid containing... or want to swap out the constant domains, the plasmids for the heavy chain and light chain are also available...ubiquitination and degradation. Find the CRBNmidi plasmid here! Kroupova, A. et al. (2024). Design of ...minimal background. Find these C. elegans mScarlet plasmids here! Cao W.X., Merritt D.M., Pe K., Cesar ...disruption in Mycobacterium abscessus using a dual-plasmid CRISPR/Cas9 system By Aryana Khosravani Deborah...Deborah Hung’s lab has developed a novel dual-plasmid CRISPR/Cas9 system for gene disruption in Mycobacterium... -
Droplet Digital PCR for AAV Quantitation
TypeBlog PostPublishedJan. 16, 2020, 2:18 p.m....some time now. When titrating AAV by qPCR, viral DNA is amplified and monitored in real time. When the...comparing the threshold fluorescence of the viral DNA to the threshold fluorescence of a standard - a set... set of reactions with known quantities of DNA. Additionally, an AAV reference, a virus with a known titer...get right. Here at Addgene, we use a linearized plasmid standard. However, even when the standard is made... the ingredients for amplification of the target DNA. This partitioning reduces the number of PCR inhibitors... circulating Mycobacterium tuberculosis-specific DNA by droplet digital PCR for vaccine evaluation in ... -
RANbodies: Reporter Nanobody Fusions
TypeBlog PostPublishedApril 10, 2018, 12:56 p.m.... projects, consider making you own DNA ladders with these plasmids from Penn State Need help picking the...are produced in animals instead of via recombinant DNA, they can’t be genetically tagged and instead are... a reporter and a nanobody that’s encoded on a plasmid. A nanobody is a ~150 amino acid, single domain... your own RANbodies? Great! All of the RANbody plasmids from Yamagata et al can be found here. Can’t find...Resources on Addgene.org More nanobody expression plasmids can be found here Watch this video to learn more... -
Tips for Titering Your Lentiviral Preps
TypeBlog PostPublishedMarch 15, 2016, 2:30 p.m....cared for your packaging cell line, prepped your DNA, transfected and harvested your lentivirus. Now it...for expression of a gene carried on the transfer plasmid or quantifying the number of viral copies that ...beneficial as they ensure that the correct transfer plasmid was used in the transfection; if preparing several... that comes from amplification of the original plasmid which can over-inflate results. A packing vector...antibiotic resistance genes carried on transfer plasmids. Target cells are transduced with serial dilutions...transduced with serial dilutions of a viral prep, genomic DNA isolated, and primers targetting viral components...In this approach, viral RNA is first converted to cDNA and then quantified using qPCR primers targeting... -
Plasmids 101: A Brief History of Plasmids and an Improved eBook!
TypeBlog PostPublishedOct. 29, 2015, 2:30 p.m....tetracycline resistant plasmid, pSC101, and a newly developed kanamycin resistant plasmid, pSC102, with EcoRI...engineering, cloning, and plasmid tags, addtional content added to the "Plasmids that Glow" and "Viral Expression...Learn more about the history of plasmids and cloning: CSHL Meeting: Plasmids: History & Biology The Joshua...Lederberg's Personal Perspective: Plasmid (1952-1997) Cohen, Stanley N. “DNA Cloning: A Personal View after...factors played within an organism. So how did plasmids get their name? In 1952, Joshua Lederberg set...inheritance factors. He proposed the catch-all term “plasmid” derived as a hybrid of "cytoplasm" and "id" (Latin...” was eventually dropped and we’ve been using "plasmid" ever since! From napkins to notebooks Although... -
Bioinformatics at Addgene
TypeBlog PostPublishedJuly 14, 2022, 1:15 p.m....scientific community obtained high-quality data for our DNA-based reagents. Not only do we extract as much information...days, we would ‘spot-check’ critical regions of plasmids deposited with us via Sanger sequencing, while...while more recently we have moved to full plasmid sequencing at our facility using Illumina MiSeq Next Generation... to a partnership with Seqera Labs. DNA sequencing drives our bioinformatics needs ...lab work as we grew. As the number of deposited plasmids increased, the accompanying sequencing data collection...transform the raw sequencing data into a complete plasmid sequence that would be ready for our QC team to...With our outline of individual steps to automate plasmid sequence assembly, and possible solutions for each... -
Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names
TypeBlog PostPublishedApril 2, 2019, 1:44 p.m.... 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..., and even humans. It binds to a specific set of DNA-binding transcription factors to repress the expression... -
Viral Vectors 101: Transductions
TypeBlog PostPublishedApril 30, 2024, 1:15 p.m....bacterial in nature. All three of these methods involve DNA incorporation into the cell, but the mechanisms vary...lentivirus, adenovirus, or AAV. Then, your desired DNA to be integrated can be cloned into the vector that...typically transfected along with viral packaging plasmids into a viral production cell line, usually HEK...HEK-293 cells. When all of these viral plasmids are expressed within the production cell line, viral particles... -
xCas9: Engineering a CRISPR Variant with PAM Flexibility
TypeBlog PostPublishedMarch 28, 2018, 6:52 p.m....transcription from the accessory plasmid (AP) encoding phage gene III. The accessory plasmid library contained all...and high DNA specificity." Nature (2018) doi: 10.1038/nature26155 PubMed PMID: 29512652 Plasmids from this...In order to bind DNA, Cas9 and other CRISPR enzymes require a short PAM sequence adjacent to the targeted...PAM flexibility. In this system, the mutagenesis plasmid (MP6) permits quick, large-scale mutation of the...Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids for Mammalian Systems Learn about CRISPR History... -
AAV Packaged on Request is Here!
TypeBlog PostPublishedJan. 10, 2025, 1:01 p.m....can request any of over 3,000 eligible AAV cargo plasmids in the Addgene repository to be packaged in your...Request With Packaged on Request, you can find your plasmids and your viral packaging service in the same place...packaged on request process, simply navigate to your plasmid of interest. If it’s eligible for packaged on request...sequencing to confirm the identity of the packaged DNA and serotype. That way, you know you’re getting the...special processes involved; simply request your plasmid of interest in your desired serotype, place your... -
AAVs CREATed for Gene Delivery to the CNS and PNS
TypeBlog PostPublishedSept. 28, 2017, 2:01 p.m....(cap) expressing plasmids is generated. Diversity is introduced into the cap plasmid library by inserting...Caltech stepped up to the challenge. Find the PHP Plasmids Here! Their Solution: Use the CREATE method to...inserting short DNA sequences that contain all possible combinations into a region of the cap gene. In this...this special CREATE cap plasmid, loxP sites flank the downstream polyA sequence. When viruses produced ...produced from this plasmid pool are injected into an animal with cell-type-specific Cre, the polyA sequence is...a cell type of interest, a scientist can isolate DNA from the infected animal and use the pre-designed... do these AAVs transduce the CNS and PNS? Both plasmids infect the PNS or the CNS much better than the... -
Your Top Requested Plasmid in 2015!
TypeBlog PostPublishedDec. 30, 2015, 3:30 p.m....from our repository to find our most requested plasmid in 2015. In line with all of the exciting developments... past year, we're excited to announce that the plasmid most requested from the Addgene repository in 2015...libraries containing thousands of pre-cloned gRNAs. DNA aliquots of the GeCKO v2 pooled libraries targeting...put to such great use. It’s just one of the many plasmids being distributed to and accelerating research...Knockouts Read Some Common Ways to Deliver CRISPR Plasmids to Mammalian Cells Resources on the Addgene Website...Visit our Lentiviral Guide Pages Find Lentiviral Plasmids Find CRISPR Pooled Libraries ... -
Your Top Requested Plasmid in 2016!
TypeBlog PostPublishedJan. 11, 2017, 8:47 p.m....for sure, as is obvious from our most requested plasmid, SpCas9 is still going strong as the basis for ...experiments. So, without further ado, the most request plasmid in 2016 was... pSpCas9(BB)-2A-Puro (PX459) V2.0... Similar to last year, the top requested plasmid from 2016 is an expression vector for CRISPR nuclease... of mammalian cell types. Purpose SpCas9 This plasmid expresses Streptococcus pyogenes (Sp) Cas9 from...sequence as directed by a gRNA. 2A-Puro In this plasmid, SpCas9 is fused to the puromycin resistance gene...resistance gene enables selection of cells with the plasmid post transfection. The 2A "self cleaving" peptide... What's not in the name? Not included in this plasmid's name are some additional features. The first and... -
5 Tips for Troubleshooting Viral Transductions
TypeBlog PostPublishedAug. 11, 2016, 2:23 p.m....hepatitis B virus DNA and hepatitis C virus RNA quantification as measured with branched-DNA technology." ...when working with virus: 1. Viral vectors undergo DNA rearrangements. Depending on the composition of ...percentage of surviving cells may be the result of plasmid integration via recombination during transfection...Recombination events during integration of transfected DNA into normal human cells."Nucleic acids research 18.9... -
Save Time and Money by Making Your Own Competent Cells
TypeBlog PostPublishedJan. 28, 2014, 3:10 p.m....survivalTM, and Stbl3TM. How to transform your plasmid DNA If you decided to use commercial competent cells...if you're transforming a plasmid with ampicillin resistance. After a plasmid is introduced to bacteria... library or an especially low concentration of plasmid), you will benefit from purchasing competent cells...negative control (plate your cells without adding plasmid) to make sure you don’t have contamination. ... Also conduct a positive control (test a plasmid that transforms well in other competent cells) to ensure...