We narrowed to 317 results for: POR C
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TypeBlog PostPublishedDec. 10, 2019, 1:58 p.m....this may be due to different lipid compositions of C. elegans membranes, which might beneficially influence...ApoB-NanoLuc reporter by inserting the NanoLuc luciferase downstream of the apoB gene. Using this reporter, the...Cell, 2019. PubMed PMID: 30639098. LipoGlo: A reporter for measuring ApoB bound lipoproteins Article ...cells. These changes were larger than previously reported for other organisms, and the Gottschalk group ...
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Plasmids 101: Inducible Promoters
TypeBlog PostPublishedJan. 18, 2018, 2:34 p.m.... Cas9 for easy genome engineering in species like C. elegans and Drosophila. Light inducible promoters... phage promoter pR, preventing expression of a reporter gene. When light is present, YFI is inactive, ...preventing repressor cI synthesis and allowing reporter gene transcription to take place. Addgene also...promoter is right for me? Here’s a list of some important variables to consider when choosing an inducible...’re expressing a gene that may be toxic, it’s important that your inducible promoter not be too leaky.... -
Building and Validating Recombinant Antibodies: A Case Study
TypeBlog PostPublishedApril 5, 2022, 1:20 p.m....Boeckman JX, Manning CF, Nguyen JT, Bechtold H, Dumitras C, Gong B, Nguyen K, van der List D, Murray KD, Engebrecht...about the process that was used to create this important collection. Creating the Plasmids To begin constructing...antibody (R-mAb) expression plasmids, allowing for important reagents that could have been lost to continue...sequences available from each plasmid webpage. Importantly, the methods developed by the Trimmer Lab for...why choosing the right secondary is of primary importance. PLoS One. 2012;7(6):e38313. doi: 10.1371/journal.pone... -
How to Deposit Your Plasmids with Addgene
TypeBlog PostPublishedFeb. 27, 2024, 2:15 p.m....depositing can be grown using standard conditions (37 °C, LB, antibiotics, etc.) and stored as glycerol stocks...drop-down menu to indicate whether the tag is on the N or C terminus and whether it was present in the backbone...be entirely verified by sequencing. Upload any support files that will make it easier for scientists to... -
Summer SciComm Series: A PhD in Science Communication
TypeBlog PostPublishedAug. 23, 2022, 1:15 p.m.... References and Resources References Drummond, C., & Fischhoff, B. (2017). Individuals with greater...how it works. I used to stay up past my bedtime, poring over my grandparents’ medical textbooks by torchlight...to all the optional sexual health sessions and reported the intel back to my shyer classmates. For my ...the community During my PhD, I’ve also found opportunities for more hands-on science communication. Between... -
Components of CRISPR/Cas9
TypeBlog PostPublishedFeb. 2, 2016, 5 p.m.... tracrRNA and Cas9, bind to these transcripts and C) cleave them leaving mature crRNAs bound to the Cas9... combined with the tracrRNA forms the scaffold portion of a gRNA and the “spacer region” forms the target... -
REPLACR Mutagenesis: Replacing In Vitro Recombination Methods
TypeBlog PostPublishedFeb. 10, 2016, 3:30 p.m.... Since the bacteria are subsequently grown at 37 °C, the recombineering plasmid is not maintained. Recombination...the deletion. REPLACR-mutagenesis successfully incorporated a 60-nucleotide insertion; beyond this insertion... -
Visualizing Protein Turnover In Situ
TypeBlog PostPublishedJan. 16, 2018, 3:20 p.m....ti2learning.org. References 1. Dieterich, Daniela C., et al. "In situ visualization and dynamics of newly...designated work areas and come with health risks. Importantly, with this method it is not possible to follow...azidohomoalanine (AHA). In the technique, AHA is incorporated into newly synthesized proteins instead of methionine...endogenous protein of interest that also had incorporated the AHA tag after pulse labeling of a determined...FUNCAT-PLA provides both quantitative and spatio-temporal information about the turnover of specific endogenous..."Puromycin inhibition of protein synthesis: incorporation of puromycin into peptide chains." Proceedings...Gabriel. "Inhibition by puromycin of amino acid incorporation into protein." Proceedings of the National Academy... -
All in a Twist: dsRNA
TypeBlog PostPublishedFeb. 2, 2023, 2:15 p.m....and References References Wang, I. X., Grunseich, C., Fox, J., et al., Human proteins that interact with... how these structures are generated, why it’s important for cells to sense their formation, and how clinicians...in turn lead to genome instability. Thus, it’s important to strike a balance between productive levels ...be delivered or anticipating this response are important considerations when using these reagents. RNA...recognized by RNase H which hydrolyzes the RNA portion, allowing the DNA strands to re-pair. The two dominating...stranded isn’t just for DNA. Untwisting RNA is important for therapeutic delivery, viruses, human disease... -
Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins
TypeBlog PostPublishedMarch 19, 2019, 1:31 p.m....Methods Mol Biol, 2018. PubMed PMID: 29043675.Grefen C and MR Blatt. Biotechniques 2012. PubMed PMID: 23066669...2017). However, Acr inhibition is spatially and temporally ubiquitous, making them a rather blunt instrument...versatility opens up the potential for spatial and temporal control in many genetic targeting and editing ...rapidly screen thousands of proteins for one that can report electrical activity via imaging. They took the ...response to light, offers a way to spatially and temporally control cellular processes in vivo. Many photoactive... -
Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur
TypeBlog PostPublishedJuly 10, 2014, 7:17 p.m....proven in (A) Productize, (B) Roll out to market, and (C) Break even cycles. Work hard. Work fast. Be flexible...bacteriophages as bacterial therapeutics. This idea was incorporated into Novophage Therapeutics, Inc. in March 2009...potential conflicts that waste money and time. Most importantly, “the company does not become real until someone...Synthetic Biology? Read On! Synthetic Biology & the Importance of Assembly Standards Story of a SynBio Startup... -
Hot Plasmids - February 2022
TypeBlog PostPublishedFeb. 22, 2022, 2:15 p.m....binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-GFP labeling various... provides a useful yet low-pressure teaching opportunity for lab members or students new to protein expression...induce high levels of gene activation for both reporter and endogenous genes. It is highly specific in... -
Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs
TypeBlog PostPublishedJune 2, 2015, 6:06 p.m....specific genome editing. Tsai SQ, Wyvekens N, Khayter C, Foden JA, Thapar V, Reyon D, Goodwin MJ, Aryee MJ...sequence flexibility indicates that gRNAs could be incorporated into these constructs without compromising cleavage...slightly higher mutation rates than they’d previously reported in protoplasts (15% and 9% of target sites.) How... -
22 Hot Plasmid Technologies from 2014
TypeBlog PostPublishedJan. 6, 2015, 5:21 p.m....targeting sequence of cytochrome c oxidase subunit VIII, so that they can be imported to the mitochondrial matrix...pCoofy series of plasmids contain a variety of N- and C-terminal tags (including His, S-tag, OneStrep, CBP...N-terminal tag, coding sequence of the gene of interest, C-terminal tag, plant terminator, and plant resistance... to report mitochondrial turnover Mitochondrial health, transport and turnover are important features...genes, reporters, or other markers into a target genome. They did this by selecting the important elements...usefulness in cell culture to report changes in mitochondrial turnover and transport. pMitoTimer from Yan’s ...Gottlieb’s lab has used an inducible MitoTimer reporter using a Tet-on system (pTRE-Tight-MitoTimer) to... -
Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**
TypeBlog PostPublishedSept. 19, 2017, 1:20 p.m....Missinato, M. A., Onuoha, E., Perkins, L. A., Watkins, S. C., . . . Bruchez, M. P. (2016). A genetically targetable...photosensitizer fluorogenic dye. MG-2I is the TAP portion of the complex and is derived from the malachite... -
Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering
TypeBlog PostPublishedMay 3, 2018, 1:48 p.m....sequence similarity to previously identified Cas13a-c orthologs, but they do include HEPN nuclease domains...localization signal (NLS). In a HEK293 mCherry reporter assay, CasRx and AdmCas13d produced 92% and 87%... -
Top Requested Lentivirus and AAV of 2016
TypeBlog PostPublishedJan. 6, 2017, 3:56 p.m....Central PMCID: PMC3785459. 2. Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, Maratos-Flier E, Roth...of transduction efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse... -
How to Keep a Lab Notebook for Bioinformatic Analyses
TypeBlog PostPublishedJune 2, 2016, 2:30 p.m....python, R, perl, and ruby // This is a comment in C++ and Java Command line history: Tracking command...your methods section about the edit that was temporarily used. Just be sure that the original code remains... -
New Tool for Lineage Tracing: The ClonTracer Library
TypeBlog PostPublishedSept. 22, 2015, 2:30 p.m....combination therapies. References 1. Hyo-eun, C. Bhang, et al. "Studying clonal dynamics in response...leukemia. Bhang says the work highlighted the importance of developing more potent ABL1 allosteric inhibitors... -
Plasmids 101: FLEx Vectors
TypeBlog PostPublishedApril 28, 2016, 2:30 p.m....References: 1. Schnütgen F, Doerflinger N, Calléja C, Wendling O, Chambon P, and Ghyselinck NB. A directional...gene. Spatio-temporal expression of optogenetic reporters: Since most optogenetic reporters are delivered...invert the reporter and GeneR fragment. Then the second recombination event would excise the reporter from ...transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic reporter would ...Cre-dependent reporter expression, the potential for background expression of the reporter will be greatly...shown in Figure 2, the cassette should contain a reporter DNA sequence (such as GFP) in the sense orientation...the two pairs of target sites. By flanking the reporter with one loxP site and one lox511 site in the ...