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We narrowed to 736 results for: RAN-1

Showing: 561 - 580 of 736 results

  1. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    ...construction of redCALWY-1, a red-shifted FRET sensor for zinc using variants of mOrange2 and mCherry as donor...field is a series of fluorescence resonance energy transfer (FRET)-based calcium biosensors. Perhaps most ...developed and published in 2010 for visualizing the transcription factor cAMP-responsive element-binding protein...really important and when? That is not known.” Transition Metals Maarten Merkx of Technische Universiteit...particularly for the intracellular imaging of transition metal ions, such as zinc, copper, and iron. “...the sensors involves plenty of trial and error. Transition metals also present challenges in that they are... way to make a FRET sensor with a large dynamic range,” he says. Merkx recently applied this “trick” to...

  2. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    ...preferentially). You can also rank the plasmids by abundance, scaled to 1, to create a “cumulative wealth... seems straightforward: E. coli bacteria are transformed with the library DNA, they grow and replicate... Each Addgene library has its own depositor transformation and amplification protocol, but the overall...Maintain representation of the library during transformation First, use enough DNA and competent bacteria...and we use electroporation to ensure a good transformation efficiency. For a typical library, Addgene ...growing the cells on solid plates, where each transformant forms its own colony, uninfluenced by the growth...concern, but Addgene still recommends plating the transformation mixture on solid media. The colonies are then...

  3. Creating accessible biology activities in schools with BioBits

    Type
    Blog Post
    .... You can email her at [email protected].   References 1. Huang, Ally, et al. "BioBits™ Explorer: A modular...that green fluorescent protein through transcription and translation. Unfortunately, having live cells in... like gel electrophoresis to separate DNA or transforming cells with green fluorescent protein, these ... into a host cell (through a process called transformation), essentially giving the cell all the instructions...enzymes that can convert certain chemicals into fragrances that they can smell, or enzymes that can create...perhaps consider using some elements of BioBits to transition your undergraduate students from the classroom...

  4. What Do I Do Now? Academic v. Non-Academic Career Decisions

    Type
    Blog Post
    ...occur, but good performers move around easily  Table 1: Taking it a Step Further - Big Company v. Small Company...encompasses millions of choices including pharma, tech transfer, management consulting, science communication,...into industry.” In his article On Leaving Academe, Terran Lane is asked by his colleagues and friends why...tell you to pick up the pace Constant worry about grants and renewals What is different about non-academia...outsource a task someone else who can do better, no grant writing Job changes can happen regularly (every ...where you want to go will help you focus on the transferable skills that you will need to get there. Additional...

  5. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

    Type
    Blog Post
    ... contain the full length plasmid.   Figure 1: The full lentiviral plasmid is expected to be >12...recombination, one resulting plasmid carries the transgene of interest (aka the viral genome) and the other...resistance gene and origin will replicate, and the transgene will be lost. Over time, bacteria containing the... grow! Our 5 Tips for Troubleshooting Viral Transductions will help with the next steps.   References...recombination of plasmid DNA without affecting mRNA translation efficiency or half-life. RNA 25:507–518. https...common lab E. coli strains Find tips for viral transduction  ...

  7. Tips for Improving Your Next Manuscript

    Type
    Blog Post
    ...class.   Four ways to improve your next manuscript 1. Be consistent with your figures The figures of a ...worthless. But simply writing down your data doesn’t guarantee a publication. There is an art to composing manuscripts...

  8. New Viral Vectors - Winter 2025

    Type
    Blog Post
    ...then scroll on for some wintery delights!    Table 1: New ready-to-use viral vectors in the Addgene repository...Request service, which offers over 3,000 eligible transfer plasmids and five different serotypes. Not sure...

  9. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    ...heterodimerizes with CIBN upon blue light exposure (see Fig 1*). When the group targeted the enhancer region of ...ubiquitin ligase, the Kanemaki lab was able to translate AID to yeast, as well as mammalian and human cell...PubMed PMID: 31026591. Expanding the targeting range of CRISPRCas9 by engineering SpCas9 Article contributed...are also thought to play a key role in the transcriptional regulation of many genes. A number of groups...thus far for these latter experiments, such as transcription factors tethered to dCas9, require long time...

  10. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    ...wide range of input DNA concentrations. This process is explained more fully in the animated Fig. 1 below...plasmids on a plate) will typically have a wide range of input quantities. This variation is problematic...produced from large numbers of samples with a wide range of input concentrations (via proprietary chemistry...differences in input concentration across a 100-fold range (1ng – 100ng) are normalized to two-fold variation...

  11. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ... and can likely be used in a wide range of bacteria. References 1. Maddalo, Danilo, et al. “In vivo...post about multiplexing with Cpf1.   Figure 1: Multiplexing allows researchers to express multiple...humanized wt Cas9, dCas9 (transcriptional repressor), and dCas9-VP64 (transcriptional activator)-containing...Multiplexing from a single transcript You can also multiplex gRNAs via a polycistronic transcript. Rather than being...being transcribed from different promoters, the gRNAs are transcribed together and are flanked by specific...simultaneously. Vectors for both transient expression and Agrobacterium-mediated transformation are available. For ...system for processing a continuous multi-gRNA transcript, each gRNA must be expressed from its own promoter...

  12. Molecular Biology Reference

    Type
    Guide
    ... DNA fragments of interest, such as genes. Figure 1: Creation of recombinant DNA. Working with Plasmids...U169 recA1 endA1 hsdR17(rk-, mk+) phoA supE44 thi-1 gyrA96 relA1 tonA EPI300 LGC Biosearch Technologies...Addgene; Promega e14-(McrA-) recA1 endA1 gyrA96 thi-1 hsdR17(rK- mK+) supE44 relA1 Δ(lac- proAB) [F traΔ36...galK2 lacY1 proA2 rpsL20 (StrR ) xyl-5 λ- leu mtl-1 Top10 Invitrogen F- mcrA Δ(mrr-hsdRMS-mcrBC) Phi80lacZM15...molecular biology, that proteins are translated from RNA, which is transcribed from DNA. The Genetic Code The..., double-stranded helix. To create the double helix, the nucleotides on the opposing strands of DNA form...proteins can be translated, the DNA is converted to RNA in a process called transcription. RNA uses the ...

  13. Plasmids 101: Origin of Replication

    Type
    Blog Post
    ...produce as many as 700 copies per cell. Figure 1: A plasmid map showing the standard features of a ...plasmids from the same group should not be co-transformed, so if you need two plasmids for an experiment... same incompatibility group should not be co-transformed. +Actual copy number varies. See below for ...stocks, agar stabs, or liquid cultures. Freshly transformed cells also give a higher plasmid yield that colonies...

  16. Antibodies 101: Epitope Tags

    Type
    Blog Post
    ...net negative charge with 4 negatively charged and 1 positively charged amino acids. Myc tags are frequently...some proteins. Protein folding issues, steric hindrances, and destabilization can be brought about by ...DOI: 10.1038/521274a Arribere, J. A., et al., Translation readthrough mitigation. Nature. (2016) 534, 719...

  17. Networking Follow-up: I Met a Person. Now What?

    Type
    Blog Post
    ...defunct (but fun) plug-in called Maps.  Figure 1: See a starfish? This is actually a map of Joanne ...enough, but I’ll never forget the strawberries. Grand gestures are memorable! Attend events for one organization... want to do: Don’t connect via LinkedIn with strangers. It’s a waste of time. People that don’t know ...

  18. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ...impartial. The best science happens when scientists have 1) the courage to make mistakes and 2) the integrity...of what I saw are that we, as a society, need to: 1) use more citations for out data so our sources are...I’ve worked tirelessly to get the publications, grant funding, and teaching experience that it takes to...

  19. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    ... lactose analogue, IPTG (isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate, IPTG binds...x-gal (5-bromo-4-chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble blue pigment...α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a functional...site (MCS) into the α-peptide (represented as an orange wedge in the figure on the left) and inserted it...produces pigment from x-gal (included in the transformation plate medium), turning the bacterial colony...IPTG is also included in the medium to ensure transcription of the lac operon.   Tips for blue-white screening...screening Use a good control: Transform the backbone plasmid without insert. All colonies on this plate...

  20. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...red light rapidly deactivates the system. Version 1 of this far-red/red light-responsive system is spread...responses are varied; one outcome is directed transcription from a given promoter. To harness the power ...the RR (OmpR) via phosphorylation, promoting transcription (see figure below). Subsequent exposure to red...response regulator via phosphorylation, promoting transcription of a GFP reporter. Subsequent exposure to red...activity persisted in the inactive state. The dynamic range, defined as the difference between the lowest and... reduced system leakiness and increased dynamic range to around 100-fold. These improvements will allow...Applications of these systems The large dynamic range of these systems ensures that they can be used with...
Showing: 561 - 580 of 736 results