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We narrowed to 373 results for: proc.2

Showing: 161 - 180 of 373 results
  1. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    Published
    Sept. 8, 2015, 2:30 p.m.
    ...Marcotte EM, Voigt CA. Nature. 2005 Nov 24;438(7067):441-2. PubMed. Find the plasmids from this publication ...Levskaya A, Voigt CA. J Mol Biol. 2011 Jan 14;405(2):315-24. doi: 10.1016/j.jmb.2010.10.038. Epub 2010...fact, precise light-based control of biological processes is highly useful in other fields, including synthetic...to directly activate and inactivate biological processes. Chemical effectors are suboptimal for a number...intensity and wavelength can be used to study dynamic processes. Conversely, varied input patterns may also be...
  2. Plasmids 101: Colony PCR

    Type
    Blog Post
    Published
    May 12, 2016, 2:30 p.m.
    ...design primers to detect the presence of your insert; 2) set up a standard PCR reaction (primers, dNTPs, polymerase...strategies for primer design: 1) insert-specific primers, 2) backbone-specific primers, and 3) orientation-specific...negative clone (-) will not. Backbone-specific primers (2) give larger sized products for positive clones (+...sequencing to verify the insert sequence before proceeding with your experiment. Shorter amplicons tend...
  3. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    Published
    April 4, 2024, 1:15 p.m.
    ... intensity of the cell’s illumination (see Figure 2). H describes the maximum signal strength, while W...SSC-A, so most people use that setting. Figure 2: As the cell passes through the laser beam and causes...scale in proportion (while H and W do not; see Figure 2), single cell events will show up in a straight line...heterogeneous cell population, for example from a processed piece of tissue, where cells are present that ...structural complexity. After several steps of tissue processing prior to running my flow analysis, I can however...interest). Let’s double-click inside our gate and proceed to the next plot! Pro tip! If you only get very...
  4. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ...the system outlined in Schmid-Burgk, J.L., et al (2). In this post, I’ll cover two great applications ...molecularly evolved from mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness...luciferin 24 hours later demonstrated a more than 2-fold brighter signal from Antares as compared to Fluc...PMID: 22894855. PubMed Central PMCID: PMC3501149. 2. Schmid, J.L., et al. (2016) CRISPaint allows modular...Here, NLuc internally excites the fluor through a process known as bioluminescent resonance energy transfer...
  5. Hot Plasmids: Summer 2025

    Type
    Blog Post
    Published
    Aug. 28, 2025, 1:15 p.m.
    ...RNA guide (known as an omega RNA, or ωRNA) (Figure 2), but TnpB is small enough to be carried by viral ...than a third of the size of Cas9). Figure 2: Comparison of TnpB (A) and Cas9 (B) genome editors...Figure 5: Mechanism of ATLASCre. VAMP2 = synaptobrevin 2; At = ALFA-tag; BACE = β-secretase; AF = AMPA.FingR...vitro and uses them in our own AAV production processes. Check out our step-by-step protocols for AAV ...
  6. Hot Plasmids: Winter 2025

    Type
    Blog Post
    Published
    March 11, 2025, 1:15 p.m.
    ...resistance to the herbicide chlorsulfuron (Homo line 2) or wild type (WT), two weeks after herbicide was ... CLIPf tag and a C-terminal SNAPf tag; see Figure 2), the authors developed a cost-effective expression... when associated with microtubules. Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET... inserted. B) Protein expression and labeling procedure. Adapted from Smith et al. 2024 under a CC-BY ...
  7. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    Published
    Oct. 27, 2015, 3 p.m.
    ...properly exiting the ER and fluorescing (see Figure 2) [5].  When using FPs containing two cysteine residues...compartments including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts...PMID: 18228466. Pubmed Central PMCID: PMC2875081. 2. Snapp, Erik Lee. "Fluorescent proteins: a cell biologist's...dark fusions is likely misfolded, blocked from proceeding through the secretory pathway, and retained in...
  8. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    Published
    April 18, 2019, 12:53 p.m.
    ...that the signal to noise ratio can be increased from 2-fold (by CRISPRainbow) to 3-fold with CRISPR-Sirius...CNV) analysis of the U2OS cells.   Figure 2: CRISPR-Sirius tricolor labeling. (A) Diagram of sgRNA-Sirius...and how these arrangements may affect nuclear processes. The latest of these visualization tools is CRISPR-Sirius...labeling of chromosomal loci in human cells.” Proceedings of the National Academy of Sciences of the United...
  9. Targeted Mutagenesis with EvolvR

    Type
    Blog Post
    Published
    Feb. 21, 2019, 1:15 p.m.
    ...while increasing the standard mutation rate by only ~2-fold over nCas9 levels. Two sets of modifications ...10-3 mutations per nucleotide per generation, and (2) the addition of the thioredoxin-binding domain (TBD... the nick site, which was expected since the processivity window of PolI is 15-20 bp. EvolvR’s on-target...bacteriophage T7 DNA polymerase increased the processivity of PolI3M, and thereby increasing the editing...
  10. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    Published
    Feb. 18, 2014, 4:22 p.m.
    ...The groups in these programs typically consist of 1-2 mentors and 3-5 mentees who meet together for an 8... together. If you all work at the same place, a 1-2 hour lunch on a regular schedule might work. If not...easier with more perspectives contributing to the process Senior people (of which there are never enough,...during the meeting.  Scientists do better with process, so don’t just get together and chat. Have an agenda...started. Have regular meetings. Experiment with the process. Have each person write a Development Plan. Set...
  11. Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC

    Type
    Blog Post
    Published
    Sept. 21, 2023, 1:15 p.m.
    ...recognize Fab fragments from the same species and 2) these fragments will be more likely to bind (and ...epitopes recognized by the secondary.   Figure 2: There are a few common solutions to avoid background...Created with biorender.com. Proper tissue processing and sample preparation can also help reduce the...
  12. Summer SciComm: Science Storytelling

    Type
    Blog Post
    Published
    Sept. 16, 2022, 1:15 p.m.
    ... (1) the struggle between two opposing forces or (2) the result of competing desires or (3) the presence... an introduction, and a resolution.   Fig. 2: The structure of a story - not just for fiction! ...find it within the science itself - within that process of failing, over and over again until you don't...
  13. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ...panning and scFv characterization.    Figure 2: Phage display starts with a pooled library of scFvs...the short epitope GCN4, also called a scaffold; and 2) a GCN4 binding scFv that’s fused to GFP. When the...Fv (Variable Domain) (scFv) Fragments. Antibodies 2:193–208 . https://doi.org/10.3390/antib2020193  Khantasup...is displayed on the coats of bacteriophages, a process which links genotype and phenotype. Incubating ...single-chain Fv analogue produced in Escherichia coli. Proceedings of the National Academy of Sciences 85:5879–5883...
  14. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ...11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016...complementary site in the genome where it will proceed to generate a blunt DNA double strand break. Cas9...stranded DNA degradation. Cas3 DNA digestion can proceed up to multiple kilobases, but only in one direction...) and does not require a tracrRNA (it can self-process pre-crRNA into crRNA). The size and simplicity ...11 cuts ssRNA and is capable of pre-crRNA self-processing. Functionally, it is very similar to Cas13 and...
  15. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...the nuclear export and translation processes as well.   Figure 2: The conserved eukaryotic polyadenylation...due to the presence of additional helper sequences (2-3). As alluded to above, termination and polyadenylation...transcriptional unit (such as a gene) and initiate the process of releasing the newly synthesized RNA from the...terminator also plays an important role in RNA processing and contributes to variability in RNA half-life...Although mostly thought of as a eukaryotic-specific process, prokaryotes also add poly(A) tails to certain ...studied as prokaryotic termination, the basic processes for eukaryotic termination are understood and ...to terminate transcription. Polymerase I uses a process similar to the prokaryotic Rho-dependent mechanism...
  16. FlipGFP, a novel fluorescence protease reporter to study apoptosis

    Type
    Blog Post
    Published
    May 21, 2019, 12:10 p.m.
    ...parts: (1) β-strands 1-9 (β1-9) and the one ɑ-helix, (2) β-strand 10 (β10), and (3) β-strand 11 (β11). Each...activating caspase-3, and detected fluorescence within 2 to 5 hours, a time consistent with previous methods...plays a pivotal role in an array of biological processes including development, the immune system, and ...cell turnover. Apoptosis is a highly controlled process that is triggered by internal and external signals...
  17. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    Published
    April 14, 2016, 2:30 p.m.
    ...be decorated with proteins and remain functional, 2) break membranes outside of their natural context ...PubMed PMID: 3759909. PubMed Central PMCID: PMC213417. 2. Schrallhammer, M. et al. Tracing the role of R-bodies...the importance of understanding these complex processes can’t be overstated, we can also learn a lot from...
  18. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ... pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays a higher...and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless...four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting ...inefficiency? Use CRISPR to make it a selectable process! As NHEJ is ineffective in bacteria, CRISPR-induced...collection of organisms, their genes, and metabolic processes, occupying the gut. Many human diseases have connections...properties of CRISPR-Cas systems is base editing. This process is generally accomplished by fusing an enzyme capable...
  19. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    Published
    Sept. 22, 2015, 2:30 p.m.
    ...represent the preexisting resistant subpopulation 2. Resistant cells among the separate replicates would...medicine 21.5 (2015): 440-448. Pubmed PMID: 25849130. 2. Hata, Aaron N., et al. "Tumor cells can follow distinct... she could “get exposure to the drug discovery process.” The ClonTracer library originally came about ...that these are selected for during the treatment process. The authors went on to show that combination treatment...
  20. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...host range plasmids. FEMS Microbiology Letters, 348(2), 87–96. https://doi.org/10.1111/1574-6968.12241  ...mobilization of broad host-range IncQ plasmids. Plasmid, 62(2), 57–70. https://doi.org/10.1016/j.plasmid.2009.05.001...negative hosts. Biotechnology and Bioengineering, 106(2), 326–332. https://doi.org/10.1002/bit.22695  Schmidhauser..., as they allow for simplified transformation processes, and can be used by scientists across disciplines...
Showing: 161 - 180 of 373 results