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We narrowed to 921 results for: Tro;

Showing: 331 - 360 of 921 results

  1. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ...enjoy seeing how our quality control process has changed with the introduction of next generation sequencing...sequencing results we analyze as part of our quality control process has steadily grown. On a busy week, we ... more than 200 plasmids as part of our quality control process. Consequently our team has refined our ...database sequence is ≥ 95%. [Our QC process would be trouble-free and much faster if 95% of our results were...Plasmids 101: An Inside Look at NGS Plasmid Quality Control Learn about our Snapgene-powered plasmid maps. ...

  2. Welcome to the Lab Bench! New Protocol Series Helps Trainees Get Started in the Lab

    Type
    Blog Post
    ...members of the lab or those taking introductory lab courses. The Intro to the Lab Bench series is meant...supplement your teaching? You’re in luck! Addgene’s Intro to the Lab Bench series was born from the shift ...meant to be an informative introduction for those who are just starting their careers in the lab and even... need a refresher on some best practices for introductory science techniques. It’s also great for teachers...

  3. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    ...capsids (i.e. the part of the virus that determines tropism) have been developed that increase transduction...expressing plasmids is generated. Diversity is introduced into the cap plasmid library by inserting short...which, in the case of Chan et al, was neurons and astrocytes. Why screen for IV deliverable AAVs? Typically... AAV vectors expressing RGB proteins under the control of an inducer, i.e. tet-off transactivator (tTA...expression in the CNS and PNS Previously to broadly introduce a transgene into a specific cell type in the brain...fluorescent tag-then-express system that allows for controlled induction of multi-color labeling of neurons ...

  4. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ...A few months ago, we shared an introduction to immunofluorescence (IF) - a common method for visualizing...of interest within a cell or tissue. In that introduction, we broke down the method into six general steps...improve antibody binding for some targets, they can destroy it for others. That said, organic solvents’ impact... K, Mareninov S, Diaz MFP, Yong WH (2018) An Introduction to Performing Immunofluorescence Staining. Methods...resources on the Addgene blog: Antibodies 101: Introduction to Immunofluorescence Antibodies 101: Epitope...Epitope Availability Antibodies 101: Introduction to Antibodies Resources on Addgene.org Find recombinant...

  5. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...researchers developed viral vector systems based on retroviruses like Moloney murine leukemia virus (MMLV). The...system that could do both, Addgene depositor Didier Trono and collaborators turned to the lentivirus HIV-1...plasmid contained a viral envelope to dictate the tropism, or types of cells the vector could infect. Finally...vectors are incredibly popular in the lab - the Trono lab’s lentiviral packaging and envelope plasmids...Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono D. Science. 1996 Apr 12;272(5259):263-7. PubMed ...Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, Naldini L. J Virol. 1998 Nov;72(11):8463-71. ...

  6. CRISPR Antimicrobials

    Type
    Blog Post
    ...resistance is swift, as few as five years after introduction. Current antibiotic strategies are nonspecific.... Phage therapy fell out of fashion with the introduction of antibiotics, but the field is currently undergoing...antibiotic-resistant bacteria (Figure 2). The first phage introduced is lysogenic, integrating into the bacterial ...lytic phage-resistant. When the lytic phage is introduced, it targets only the bacteria that are antibiotic...gain resistance to a lytic phage through the introduction of a crRNA (shown in blue). Infection with the...Bacterial Genome Engineering Treating Muscular Dystrophy with CRISPR Gene Editing Using CRISPR to Edit ...

  7. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...unable to reach all regions of the CNS. PHP.B, a neurotropic variant of AAV9, is better than AAV9 at gaining...receptor used by PHP.B viruses. Creating PHP.B, a neurotropic AAV9 variant PHP.B’s specificity for C57BL/6J...CNS transduction and transduces the majority of astrocytes and neurons across multiple CNS regions. PHP....Ly6a seemed the most likely candidate due to its strong linkage to the PHP.B phenotype and its expression...mice strain specific Ly6a SNPs on PHP.B binding in vitro. They found that when LY6A contained C57BL/6J SNPs...PMID: 30819613. Hordeaux, Juliette, et al. "The neurotropic properties of AAV-PHP. B are limited to C57BL...

  8. Hot Plasmids - August 2020

    Type
    Blog Post
    ...! The team used other molecular techniques to introduce time modulation of the TEV cleavage and improve...dCas-KRAB activator and the dCas-VPR repressor are introduced into the hPSC genome using AAV1 and contain an...Find the plasmids at Addgene! CRISPR RNP electroporation and AAV donor infection (CRISPR-READI) enables...new tool for synthetic biology uses CRISPRi to control gene expression. This includes a CRISPRi-based ...affinity), as well as GRAB_NEmut as a negative control. Find the sensors at Addgene! New yellow fluorescence...AAV5 and AAVrg. Find these AAVs at Addgene New controls, biosensor, chemogenetic and optogenetic AAV for...

  9. Hot Plasmids - November 2023

    Type
    Blog Post
    ...New compact, efficient prime editors By Susanna Stroik The Liu lab recently used phage-assisted continuous...limitations and enable impressive editing efficiencies in vitro and in vivo. Incorporating reverse transcriptase... vectors, CreVLPs, and eVLPs for in vivo and in vitro applications.   Figure 2: Modular features... enzyme, Lee and Cheah et al. achieved precise control over when and where the enzyme is active, drastically..., then the other, a dual-enrichment and mass spectrometry workflow identifies proteins bearing both labels...light-regulated LOV-Turbo enables precise spatiotemporal control of proximity labeling in living cells. Nature Methods...

  10. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    ... it is essential to include proper controls. As a positive control, include a sample that is known to ...This method also requires more optimization and controls when developing, as secondary antibodies can also..., purified protein, or a peptide. The positive control will confirm that the procedure is working and ...and should always be included.  For a negative control, include a sample that does not contain the target...where the target is not expressed. The negative control will help you determine if there is non-specific...

  11. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ...size using electrophoresis, typically capillary electrophoresis. Capillary electrophoresis reads a fluorescent...process, see our blog post on NGS plasmid quality control. NGS is generally cheaper if you have many samples...to sequence our packaged viral DNA for quality control. This gives us a faster turnaround time than short-read...Validation: seqWell Addgene’s Tips for Plasmid Quality Control Additional Resources on addgene.org Molecular...

  12. The Challenges of Cell Culture

    Type
    Blog Post
    ...most as cancer cells are amenable to culture in vitro. Virologists too have benefited from the ability... ‘Master’ stock, which can be stored in liquid nitrogen long term, preferably some of these vials will...will be stored in a separate backup liquid nitrogen store to prevent loss from equipment malfunctions. One...also monitor the stock levels you have in liquid nitrogen for your Working and Master stocks of each cell... laboratories to carry our independent quality control for a range of testing, from animal doping to forensics...Thermofisher Guidelines for Maintaining Cultured Cells Invitrogen Cell Culture Basics Guide Abcam Cell Culture ...

  13. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...are available to confer greater transcriptional control, assist with proper protein folding, and deal with...Contains mutated lac permease whch allows for linear control of expression F- ompT lon hsdSB (rB- mB-) gal ...polymerase (RNAP) from the bacterial genome under control of the lac repressor, which is inducible by the...cases, some strains carry an additional measure of control such as the pLys plasmid, which suppresses basal...difference being that the latter provides tighter control of basal expression. What if I don't see protein...sources, so fear not—there are a few simple troubleshooting measures that can help get you back on track...

  14. Enabling high school research at the Journal of Emerging Investigators

    Type
    Blog Post
    ...Emerging Investigators Podcast breakdown Intro Chapter 1 - Introduction to Mark Springel, Olivia Ho-Shing, ... In this episode of the Addgene Podcast, we introduce you to the Journal of Emerging Investigators, an...developing one-on-one mentorship programs. Chapter 6 and Outro - Interview with high school student researchers...basic plasmid biology Use or CRISPR 101 eBook to introduce students to this amazing genome enegineering technology...

  15. New Podcast Segment: Hot Plasmids

    Type
    Blog Post
    ...Yeast Prototrophy Kit from Markus Ralser's Lab -  Use this kit to complement unused auxotrophies in Saccharomyces...to a quick (~5 min) Hot Plasmids podcast that introduces you to 4 new plasmid technologies from one of... Plasmid 2: Magneto 2.0 form Ali Guler's Lab - Control neuronal activity with a magnetic field. Listen...strains.  Listen to Our Segment on The Yeast Prototrophy Kit Special thanks to our sound engineer/producer...

  16. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ..., such as the introduction of heterozygous endogenous epitope tags, or the introduction of dominant mutations... examples from their lab include using AAV to introduce epitope tags into the endogenous alleles of the...2008). They have also used AAV gene editing to introduce naturally occurring cancer-causing mutations into...However, while CRISPR is remarkably effective for introducing non-sequence specific frameshift mutations through...efficiency of AAV-based gene editing via the introduction of a double strand break by using a targeted...

  17. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...coverage, mutational efficiency, control over where mutations are introduced, and scalability (see Supplementary...favoring of certain types of mutations, difficulty controlling the rate of mutagenesis, and low throughput. ...mutagenesis PCR that uses mutagenic primers to introduce all desired codon changes (step 1 in the image... high-fidelity Phusion polymerase are used to introduce point mutations into the newly synthesized DNA...Error-Prone PCR”. Methods in Molecular Biology In Vitro Mutagenesis Protocols, pg. 103-109. PubMed PMID:...

  18. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...Shine-Delgarno sequences used in this so-called bicistronic design (BCD). The 49 vectors in this kit can ...of varying strengths centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013...terminators can be used to precisely tune your polycistronic E. coli expression constructs. Some applications... Living Bacteria Using an Orthogonal Ribosome-Controlled Fluorescent Reporter."ACS chemical biology 11.1...vector that enables protein expression in any in vitro translation system. The gene set was carefully chosen...been used to produced recombinant proteins for in-vitro assays: Koch, Daniel, et al. "A pull-down procedure...

  19. Evolution of Lab Techniques

    Type
    Blog Post
    ... the strength of the signal in comparison to a control sample. Southern blotting applications range from...causes sickle cell anemia. The mutation happens to destroy a sequence recognized by the restriction enzyme...gels and next-generation sequencing techniques Introduced in 1977, Sanger sequencing, also known as chain-terminating...referred to as sequence DNA as it synthesized by electronically recording fluorescence activity as fluorescently...differences in ion flow caused by the different electrochemical properties of nucleotides (8). Compared to ...sequences among DNA fragments separated by gel electrophoresis." Journal of molecular biology98.3 (1975): ...

  20. Save Time and Money by Making Your Own Competent Cells

    Type
    Blog Post
    ... competent cells (protocol) Preparation of electrocompetent cells (protocol) At Addgene, we use the...competent cells (protocol)  Transformation of electrocompetent cells (protocol)  Cells prepared using ...with ampicillin resistance.  After a plasmid is introduced to bacteria, the bacteria is typically grown ...competent cells, don’t forget to do a negative control (plate your cells without adding plasmid) to make...have contamination.  Also conduct a positive control (test a plasmid that transforms well in other competent...

  21. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    ...A few years ago, researchers introduced what’s been called a major breakthrough in cloning technology...plant model systems. “The cloning is all done in vitro; it’s no different in mouse, human or plants,” explained...of an advanced molecular plant science lab.” Introducing GreenGate As Lohmann and his colleagues describe...describe in the PLoS ONE paper that introduced the kit in December, GreenGate includes the six most frequently... available through Addgene,” Lohmann said. “We strongly believe the free sharing of material is absolutely...

  22. Six Spooky Science Stories and Halloween at Addgene

    Type
    Blog Post
    ...and did their best to claim the costume contest trophy. We’ve had teams dressed up as plasmids, Blugene...unilateralis is a fungus that infects ants and controls their behavior. These ants become “zombies” and...their jaws into a leaf, and dies as its muscles atrophy. After the ant’s death, the fungal fruiting bodies...display imitating a moving caterpillar. The worm controls the snail’s behavior, forcing it out of hiding...blood out. Once the blood is depleted, the tongue atrophies and falls off. The louse remains becoming the ...

  23. A new platform for fast functional characterization of potentially cancerous mutations

    Type
    Blog Post
    ... in a pooled cell population, as mutations with strong phenotypic effects can mask weaker ones. Thus, ... the need for a faster and higher-throughput in vitro approach. Using data from The Cancer Genome Atlas...them had not yet been functionally validated. In vitro assay allows first-time functional annotation of...collection to screen individual perturbations in vitro using Ba/F3 and MCF10A cellular models. Ba/F3 and...effect on cell viability compared to a wild-type control. Approximately ~22% of the mutations, however, ...

  24. Trimmer Lab NeuroMab Collection

    Type
    Collection
    ...undergo application-specific validation and quality control by Addgene as well as by our trusted partner labs...

  25. Supporting Rare Disease Research with BeHeard 2017

    Type
    Blog Post
    ...afflictions. Elia Di Schiavi - Spinal muscular atrophy Dr. Elia Di Schiavi is a Research Scientist at ...Naples, Italy. His group works on spinal muscular atrophy, a disease in which disruption of the smn1 gene...the smn1 gene is known to cause spinal muscular atrophy, the molecular mechanisms behind the disease are...restore motor neuron function in spinal muscular atrophy patients. Noushin Nabavi - Peritoneal Mesothelioma...of incidence and prevalence of spinal muscular atrophy (SMA)." European Journal of Paediatric Neurology...

  26. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ... The results are then visualized using gel electrophoresis. Surveyor™ and T7E1 have been widely adopted...research setting, but these events could have disastrous health effects in the clinic. NGS-based methods...including Digenome-seq (Kim et al. 2015) for in vitro analysis, IDLV for in vivo detection (Gabriel et...potentially toxic) Digenome Seq 0.1% Cell-free (in vitro) Works across all cell types Must be verified with... background BLESS Not reported Cell-based (in vitro) Can be used on tissue from whole animal models,...most investigators use single cell clones for in vitro CRISPR experiments. The likelihood that a single...

  27. CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

    Type
    Blog Post
    ...of input material, cells or tissue, to produce a strong enough signal over background noise. This is not...access to transcriptional regulatory elements is controlled during development using the Drosophila wing ...the target protein. This targeted digestion is controlled by the addition of calcium, which MNase requires...only for a short period of time, thus precisely controlling the amount of cutting and mitigating noise generated...protocol, the most significant being the usage of a strong detergent to permeabilize cells rather than to ...

  28. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    .... Layers of control have also been added to the CRISPR technology to allow time-controlled edits to occur...target can be identified if it isn’t known. If a strong candidate emerges, downstream optimization is performed...generation of knockout cell lines which has made this control step extremely accessible. For example, let’s say...  Susanna Stroik...

  29. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    ...traps can be used as high-throughput screens to introduce reporters throughout an organism’s genome. These...Enhancer traps do not need to be integrated within an intron or exon of a gene, and thus they do not disrupt...splice acceptors. They require insertion into an intron of a gene, where they can be spliced into the gene...Additional resources on Addgene.org Vector database Intro to the Lab Bench Plasmids 101 e-book Additional... Susanna Stroik...

  30. Networking and Inspiring Young Scientists at Freshers Fairs

    Type
    Blog Post
    ...ways budding young scholars across Europe get introduced to the cultures and opportunities available to... societies to promote their upcoming events and intro sessions, and give out useful items and information...fairs are also where many future scientists are introduced to the biology and chemistry societies that will... to attend. We thought we’d take some time to introduce you to the idea of the Freshers Fair and some ...Scientific societies like those mentioned above provide strong foundations upon which young scientists can build...
Showing: 331 - 360 of 921 results