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We narrowed to 605 results for: RAN-1

Showing: 241 - 250 of 605 results
  1. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    Published
    Aug. 15, 2019, 12:30 p.m.
    ... requires selection. These two steps are (Figure 1): The initial plasmid integration step (the so-called...antibiotic resistance marker from the vector (abR, Figure 1). The resolution step (“double crossover”) which leads... loss of a specific phenotype (using sacB, Figure 1, further discussed below).  Counter-selectable marker...Current protocols in molecular biology 106.1 (2014): 1-16. PubMed PMID: 24733238. Additional resources on...strong transcriptional terminator BBa_B1002 (from IGEM) upstream of the MCS to reduce transcriptional readthrough...rely on exogenous enzymes, require multiple transformation and plasmid curing steps, and can generate ...worked too. Rhamnose induction relies on the transcriptional activator rhaS. To allow manipulation of bacteria...
  2. Transferable Skills Guide: Creativity

    Type
    Blog Post
    Published
    Jan. 18, 2022, 2:15 p.m.
    ...yourself in a creative rut, read on!   Fig 1: Blugene using creative problem-solving in the lab...Blog Transferable Skills Guide: Problem-Solving Transferable Skills Guide: Teamwork Transferable Skills...feel they lack formal training in a variety of transferable skills. At Addgene we're helping to fill this...both helping researchers recognize their MANY transferable skills learned while working in the lab and ...Today, we’re highlighting an oft-underlooked transferable skill: creativity. I’ve heard quite a few scientists..., it’s clear that creativity is absolutely a transferable skill developed during PhD training. Creativity...problem. In a study outlined in Originals by Adam Grant, a creativity expert at Stanford conducted an experiment...
  3. Human Germline Editing Using CRISPR

    Type
    Blog Post
    Published
    Aug. 10, 2017, 2:19 p.m.
    ... employing HDR. Ma et al. moved forward with gRNA 1 to conduct microinjection experiments in S phase zygotes...instead from the mother’s wild type allele (Figure 1). 72.4% of embryos carried two wild-type copies of...proceeded through NHEJ rather than HDR. Figure 1: Homology-directed repair in targeted zygotes proceeds...the embryos underwent repair not from the single-stranded oligonucleotide (ssODN) template supplied by Ma...maternal allele rather than the supplied single-stranded oligonucleotide (ssODN) template. Embryos also...off-target effects, but this advantage likely won’t transfer to all genes we’d like to target. In addition,...
  4. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ...succumb to the toxicity of CcdB.   Figure 1: Image from Bernard, P., et al. showing the pKIL 18...submit a guest blog entry of your own! References 1. Bernard, P. “Positive Selection of Recombinant DNA... plasmid ccdB killer gene”. Gene. 1994 Oct 11;148(1):71-4. PubMed PMID: 7926841. 4. Bernard, P., et al...poison, locking up DNA gyrase with broken double stranded DNA and ultimately causing cell death. ccdA, another...gene in frame with an MCS. E. coli that were transformed with the empty vectors expressed the ccdB gene... that ccdB Survival™ can be more difficult to transform, depending upon the specific plasmids being used...
  5. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...destabilize the transcription complex, initiating cleavage of the transcript.   Figure 1: A predicted... monkey virus constitutive RNA transport element lead to enhanced HIV-1 Gag expression in a context-dependent...prokaryotic terminators and deposited several with Addgene (1). Prokaryotic polyadenylation Although mostly thought...initiation step of gene transcription; today we'll provide an overview on how transcription stops, or termination...from the transcription machinery. Terminators are found downstream of the gene to be transcribed, and typically...generally serves to promote transcript longevity in eukaryotes and promote transcript degradation in prokaryotes...Hepatitis Virus Posttranscriptional Regulatory Element Enhances Expression of Transgenes  Delivered by ...
  6. Michael Koeris' Journey from Grad Student to Entrepreneur: The Story of Sample 6

    Type
    Blog Post
    Published
    Jan. 21, 2016, 4:52 p.m.
    ...Podcast breakdown (Time : Topic) 0:00 - 1:25 : Introduction 1:25 - 6:19 : Dr. Koeris' graduate work in...Novophage's successful transformation into Sample 6 and Dr. Koeris' own transition from graduate student...37-36:53 : Was It difficult for Dr. Koeris to transition from developing therapeutics to developing a ...
  7. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    Published
    March 12, 2020, 1:15 p.m.
    ... DNA that were transcriptionally active in retinal cell types in mice. Figure 1: The four strategies...used four different design strategies: Strategy #1 (ProA): Create promoters with sequences upstream of...highest rate of targeted expression. Surprisingly, <1% of synthetic promoters replicated the expression ...mouse cell type specific transcription factor binding sites interspaced with random sequences.   Strategy...synthetic promoters. Legend: TSS: transcription start site. TFBS: transcription factor binding site. Image from...particular cell type is important not only for translational research, such as gene therapy, but also basic... using naturally occurring promoters to drive transgene expression, the Roska lab built synthetic promoters...
  8. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    Published
    April 19, 2018, 1:25 p.m.
    ...! References 1. Sinche, Melanie, et al. "An evidence-based evaluation of transferrable skills and job...Page Read Other Posts in the Transferable Skills Guide Developing Transferable Skills During Science Training...that they lack formal training in a variety of transferable skills. At Addgene we've set out to fill this... highlighting that researchers do learn MANY transferable skills while working in the lab and by offering...where you might need some help. Today in our transferable skills guide: Collaborating with others outside... had to overcome some hesitation to ‘approach strangers’ and learn how to communicate outside my scientific...message and to keep it short. Soon, discussing with strangers stopped feeling weird and started to be fun. I...
  9. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    Published
    April 12, 2016, 2:30 p.m.
    ..., including those found at Addgene! Figure 1: Strategy for assembling the barcoded gRNA library...pool-clone them into the pAWp28 storage vector (Figure 1): Forward oligo: 5’- CACCGNNNNNNNNNNNNNNNNNNNGTTTGGGTCTTCGAGAAGACCTATTCXXXXXXXXC... He can be contacted at [email protected].   References 1. Cheng, Allen A., Huiming Ding, and Timothy K. Lu....expression of other genetic elements including transcription factors (Cheng et al., 2014) and microRNAs (.... Konermann, Silvana, et al. "Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex...
  10. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ...various cyan fluorescent proteins is depicted in figure 1. Of note, the photobleaching rate does not depend ...to be used with the fluorescent protein. In figure 1 we provide an example of such a comparison between...the variants is comparable (lower panel of figure 1). To avoid changes in excitation power, it is important...fluorescent proteins in the same spectral class (Cranfill et al, 2016). This non-linear dependence on excitation...the conclusions reached at high power may not translate to real applications in which far less power is...
Showing: 241 - 250 of 605 results