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Showing: 61 - 80 of 101 results
  1. Your Top Requested Plasmid in 2015!

    Type
    Blog Post
    Published
    Dec. 30, 2015, 3:30 p.m.
    ...PMID: 24336571. PubMed Central PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang...metalloproteinase-9 secretion, and identifying lysine methyltransferase substrates. We’re incredibly happy that lentiCRISPR...
  2. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    Published
    Sept. 17, 2024, 1:15 p.m.
    ... that all look exactly alike.      Figure 2: Reagent gradient. Here, the lanes are lightly marked...guide. The gel is then removed from the membrane (2) and the membrane is cut so each lane is its own piece...in the bands, you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to ...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ...incubation (10–20 minutes) at 70 °C, or a truly lengthy incubation (30–60 minutes) at 37 °C, if you suspect...
  3. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    Published
    Nov. 10, 2016, 3:30 p.m.
    ...diffusion test or Kirby-Bauer susceptibility test (Figure 2), and requires at least 18-24 hours to obtain results...." London: Wellcome Trust & HM Government (2016). 2. CDC Information Pages on Antibiotics 3. WHO Antimicrobial...flu) Overuse of antibiotics in livestock farming Lengthy courses of treatment with antibiotics These scenarios...
  4. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked by two direct...nucleotide sequences. To counter this, methyltransferases add methyl groups to bases at specific nucleotide...more stable than the methyltransferase. When a plasmid is lost, the methyltransferase is degraded and/or...composed of restriction endonucleases and methyltransferases. Restriction endonucleases cleave DNA at ...the cell grows resulting in the appearance of unmethylated sites in the chromosome. The more stable endonuclease...
  5. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ...cryptochrome 2 are the chromophores flavin-adenine dinucleotide and methyltetrahydrofolate, which are universally...
  6. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    Published
    April 4, 2023, 1 p.m.
    ...work with Grows extremely fast (doubling time of ~ 2 hours) Amenable to genetic manipulation The first...operates as an obligate aerobe. It’s also a methylotropic yeast, meaning it can utilize methanol as its...peroxisomes are where methanol metabolism occurs in methylotrophic organisms. K. phaffii is an excellent model...
  7. CRISPR in the Clinic

    Type
    Blog Post
    Published
    Nov. 2, 2023, 1:15 p.m.
    ... Cure for HIV/AIDS. Int J Mol Sci. 2023 Jan 13;24(2):1563. doi: 10.3390/ijms24021563 Resources on Addgene.org.... Finally, allogenic therapies involve editing healthy donor-derived cells and delivering them to the ... have been modified ex vivo, previously edited healthy donor cells can be delivered immediately, saving...clinical trial for SCD in 2022 as well. Hereditary transthyretin amyloidosis (hATTR) hATTR is caused by a variety...
  8. Top 10 Open Science Developments of 2013

    Type
    Blog Post
    Published
    Dec. 19, 2013, 3:46 p.m.
    ...access to the results of federally-funded research. 2)      In June, the White House honored 13 “Champions... a few minutes to reflect on some of the most noteworthy developments in open science this year. Here ...
  9. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    Published
    July 30, 2024, 1:15 p.m.
    ...’re delivering their payload to.   Figure 2: Numbers of AAV tools, by category, available from... substrates, light, or engineered ligands (Figure 2). Biosensors, which include calcium and voltage sensors...significant time and removes a variable from the lengthy process of validating and using an AAV in a specific...
  10. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    Published
    June 20, 2024, 1:15 p.m.
    ...the chromosome, ending replication.   Figure 2: Prokaryotic chromosomal replication. Adapted from...Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/MMBR.62.2.434-464.1998...contribute to this regulation in different ways: methylating newly synthesized DNA, blocking binding sites...
  11. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ...Central PMCID: PMC323600. 2. Chester, Nicholas, and Daniel R. Marshak. "Dimethyl sulfoxide-mediated primer...isolated from a methylation competent bacterial strain (e.g. DH5α which is dam+). Methylation deficient bacterial...template. The parent template is removed using a methylation-dependent endonuclease (i.e. DpnI), and bacteria... of the primers. Because you will be using a methylation-dependent enzyme (DpnI) to eliminate the parent...is unique in that it cleaves only DNA that is methylated at the adenosine of the GATC recognition site...
  12. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...restrict methylated DNA fhuA2 lacZ::T7 gene1 [lon] ompT gal sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R... restriction/methylation system. This means the strain can neither restrict nor methylate DNA. dcm: Similarly...Similarly, strains with this mutation are unable to methylate cytosine within a particular sequence. Browse... in the exponential growth phase are alive and healthy, which makes them ideal for protein expression....
  13. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...of concept for further development. Figure 2: Mechanisms of deaminase-based adenine transversion...hypoxanthine) or between the pyrimidine bases (cytosine, thymine, and uracil). Base transversions are edits that...have an enzyme to remove inosine from DNA: N-methylpurine DNA glycosylase (MPG), also known as alkyladenine...BioRender.com. Base excision editing extends to thymine Since an enzyme that excises inosine can be modified...which excises uracil, could allow it to recognize thymine (T) or cytosine (C). Several different teams succeeded... DNA, creating an abasic site. MPG / AAG N-methylpurine DNA glycosylase / alkyladenine DNA glycosylase...editing by fusing an adenine base editor with an N-methylpurine DNA glycosylase. Nature Biotechnology, 41(8)...
  14. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ... from damage (Abe & Lim, 2024).    Figure 2: A) When proteins (blue) interact with the air-water...editor consisting of dCas9 fused to a DNA methyltransferase — that can drive lasting transcriptional silencing...Histone tail for Autoinhibition Release of Methyltransferase (CHARM), with several advantages over the ...  Rather than overexpressing an exogenous methyltransferase for epigenetic silencing, which can be toxic...tail to recruit and stimulate the endogenous methyltransferase. The strategy is generalizable to a variety...D3L recruits the endogenous, autoinhibited methyltransferase DNMT3A to the genomic location specified by...DNA-binding protein. The ADD domain of the methyltransferase binds to H3K4me0 on D3L, releasing the autoinhibition...
  15. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    Published
    July 12, 2016, 2:30 p.m.
    ...posts: Part 1: Genetically Engineered Mice and Part 2: Breeding and Crossing Mice. My first CRISPR mouse...is less predictable. In our best case, we saw that 2 out of 3 mice carried a 5 kb insertion in the ROSA...:1262–1267 . https://doi.org/10.1038/nbt.3026 Gurumurthy CB, Takahashi G, Wada K, Miura H, Sato M, Ohtsuka...
  16. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    Published
    Jan. 4, 2022, 2:15 p.m.
    ...become a valuable asset yourself.     Fig 2: A diagram of the Thomas-Kilmann Conflict Model, courtesy...as well. What I really brought to the table was empathy, a willingness to listen, and an ability to remember... are a few more tools that can come in handy: Empathy: If you often find yourself in confrontational ...critical component of conflict resolution. Having empathy does not mean you like everyone you meet. Most ...solid examples of times when you have used your empathy, listening skills, and institutional knowledge ...
  17. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    Published
    Oct. 8, 2020, 1:15 p.m.
    ...bringing in environmental contaminants.  Figure 2: An Addgenie works in the tissue culture hood while...doi.org/10.1290/1071-2690(2002)038<0086:caeomi>2.0.co;2 Uphoff CC, Drexler HG (2011) Detecting Mycoplasma ...increases the risk of secondary contamination to other healthy cell lines. Mycoplasma Removal Agents (MRA) are...
  18. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    Published
    Feb. 13, 2025, 2:15 p.m.
    ... a template, copying the base edit. Figure 2: Cytidine deamination takes place on the free strand...SpCas9 base editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated...formation. To further decrease indel formation 1.5-2 fold, the team fused bacteriophage protein Gam to ... DNA. Uracil is then subsequently converted to thymine through DNA replication or repair. Fusing an inhibitor...
  19. Plasmids 101: Gram Negative and Positive Bacteria

    Type
    Blog Post
    Published
    Aug. 2, 2022, 1:15 p.m.
    ...Bacterial Cell Envelope. Cold Spring Harb Perspect Biol. 2(5) a000414 (2010). Cleveland Clinic. Gram Stain. https...iodine solution. Rinse briefly with tap water. Add ethyl alcohol or acetone drop by drop onto the smear until...
  20. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    Published
    Aug. 23, 2022, 1:15 p.m.
    ...perceived climate change risks. Nature Climate Change, 2(10), 732-735. doi:10.1038/nclimate1547 Kunda, Z. (...were much more likely to rate that study as untrustworthy, inaccurate, methodologically flawed, and biased...
Showing: 61 - 80 of 101 results