Skip to main content
Addgene
Showing: 261 - 274 of 274 results
  1. MXS Chaining

    Type
    Blog Post
    Published
    Feb. 7, 2017, 3:30 p.m.
    ...20205762. PubMed Central PMCID: PMC2822740. 2. Engler C, Kandzia R, Marillonnet S. A one pot, one step, precision...CpG dinucleotides in their 6-mer sequences, are rarely found in the transcriptomes of these species and...
  2. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ...Zou, Y., Mason, M.G., Wang, Y., Wee, E.J., Turni, C., Blackall, P.J., Trau, M., & Botella, J.R. (2017)...around. Finally, purified total nucleic acids are released by dipping the cellulose directly into a PCR buffer...
  3. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    Published
    April 3, 2014, 8:05 p.m.
    ... mRNA Mammalian promoter from the human ubiquitin C gene Constitutive  As the name implies, this promoter...cells and typically only in the same or closely related species from which they were derived. Similarly...
  4. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ..., E. O., Williams, B. A., Zhang, J., & Chaput, J. C. (2010). “Random Mutagenesis by Error-Prone PCR”. ... protein of interest’s function. This technique relies on the generation of a plasmid library that expresses...mutagenesis (SDM) PCR, but with a twist since it relies on the sequential nicking, degradation, and PCR-generation...
  5. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...2.50021-0  Toh, W. K., Teo, Y. L., Tor, X. Y., Loh, P. C., & Wong, H. L. (2023). Development of constitutive...aurum, M. smegmatis, Paracoccus denitrificans, Pasteurella multocida, Porphyromonas gingivalis, Proteus ...
  6. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    Published
    April 4, 2024, 1:15 p.m.
    ...in fewer cells being considered for the analysis. (C) A more liberal gate allows more cells to be considered...looser gate here. I am not worried about including irrelevant events in my analysis, as these will get excluded...from your single cells, as duplicates can’t be reliably analyzed on their staining signals. For this, ...    Figure 4: (A) Using the proportional relationship between FSC-H and FSC-A we can gate on single...displaying your events. You are free to choose any size-related parameter, such as FSC-H, FSC-A, SSC-H, SSC-A, ...Figure 6: (A) A flow plot showing two stains in relation to each other. Using a quadrant gate, I can divide...
  7. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    Published
    Aug. 15, 2019, 12:30 p.m.
    ...2015): 15096. PubMed PMID: 28060411. Thomason, Lynn C., et al. "Recombineering: genetic engineering in bacteria...methods roughly fall into two categories: those that rely on exogenous enzymes and those that use endogenous...). Both of these systems are powerful, but they rely on exogenous enzymes, require multiple transformation...The most commonly used counter-selection system relies on the sacB gene product of B. subtilis (Hmelo ...would allow stronger counter-selection than one merely co-opted toward that aim (i.e. sacB). Importantly...arabinose promoter worked too. Rhamnose induction relies on the transcriptional activator rhaS. To allow...
  8. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    Published
    Feb. 8, 2024, 2:15 p.m.
    ...3):937-948. doi: 10.1016/j.ymthe.2020.11.009  Guo C, Ma X, Gao F, Guo Y. Off-target effects in CRISPR/...is low, but since all downstream experiments will rely on that clone not having any confounding mutations...sequencing The only way to quantify off-target effects entirely is to perform whole genome sequencing. While this...major con is that binding by Cas does not always correlate with catalytic activity at those sites, especially...and integrase-defective lentiviral vectors - which rely on insertion of DNA by the NHEJ pathway during DNA...
  9. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    Published
    Dec. 6, 2022, 2:15 p.m.
    ...a variety of imaging conditions (155 μs and <0.1 °C in optimal conditions). While B-gTEMP itself is a ... lab, Kaspar Podgorski and colleagues have now released iGluSnFR3 (Aggarwal et al. 2022). This new version...measurement of different RTKs. Although this sensor relies on a change in localization to the cell membrane...
  10. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ... easy for a researcher to quickly generate N- and C-terminal epitope tag fusions to a protein of interest... was carefully chosen to perform statistically relevant benchmarking of cell-free expression systems, ... the efficiency of their cell-free system in correlation to the size of the product. Analysis of protein...
  11. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    Published
    May 18, 2018, 7:32 p.m.
    ...repository 49:45 0 1:18:45 - Reagent Development - Steven C. Almo, Institute for Protein Innovation  1:18:45 -...45), there are also initiatives to create more reliable reagents for future research. I’ve talked to many...many researchers who have complaints about the unreliability of antibodies. IPI is working to address this...repeat previously published results. This over-reliance on the perfect story for a “high impact” publication...ways to publicise negative data - we don’t have to rely on traditional journals. There’s social media - ...publishing community views reproducibility, how industry relies on reproducibility, and how you can push for more...
  12. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ...composed of Venus fluorescent protein, the Troponin C Ca++ binding domain, and NanoLuc® Luciferase. CalFlux...Many intracellular sensors of events like calcium release have been made using fluorescent resonance energy...biosensors, the intensity of the FRET signal can be correlated to specific concentrations of biological molecules...et al. (7) wished to trigger intracellular Ca++ release with an optogenetic receptor, melanopsin, requiring...
  13. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ... Y, Miki T, Fujioka R, Uehara R, Tomioka A, Obuse C, Kubo M, Hiwatashi Y, Goshima G (2012) An Inducible... of CRISPR technologies, one might incorrectly relegate RNAi to history. The straightforwardness of RNAi...2018) and the model bryophyte Physcomitrium (Physcomitrella) patens (Bezanilla et al., 2005; Nakaoka et...Klueh P, Quatrano RS (2005) An RNAi System in Physcomitrella patens with an Internal Marker for Silencing...Inducible RNA Interference System in Physcomitrella patens Reveals a Dominant Role of Augmin in Phragmoplast...
Showing: 261 - 274 of 274 results