We narrowed to 856 results for: TOR
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TypeBlog PostPublishedMay 26, 2016, 2:30 p.m.... like PCR, cloning, transfection, or long-term storage. But what about those pesky situations where your...maximize cell density and plasmid yield as these factors are generally, but not perfectly, correlated. ...manufacturer. On Addgene plasmid pages, you may see depositor recommendations to grow a plasmid at 30℃ or room...
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Genome-wide Screening Using CRISPR
TypeBlog PostPublishedAug. 18, 2015, 2:30 p.m....population of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within the genome...gRNAs are then cloned into lentiviral transfer vectors to create the CRISPR library. CRISPR libraries ...library that is right for you There are several factors to consider when selecting a CRISPR library for... -
Tips for Using BLAST to Verify Plasmids
TypeBlog PostPublishedMay 29, 2014, 1:29 p.m....the sequencing results of the plasmids in our repository with known reference sequences, such as full ...full plasmid sequences provided by the laboratories that deposit their plasmids with us or other entries ...entries in NCBI’s numerous databases. As our repository has grown over the years (we now have over 60,000 plasmids... -
Which Fluorescent Protein Should I Use?
TypeBlog PostPublishedMay 20, 2014, 2:06 p.m....Protein (GFP), a protein from the jellyfish Aequorea Victoria. Since that discovery, the family of FPs just ... used are GECIs - genetically encoded calcium indicators (e.g. GCaMP). Others include: pHluorin & pHTomato...have a filter that will pass blue light to the detector/camera, then BFPs are of no use to you. When using... -
MXS Chaining
TypeBlog PostPublishedFeb. 7, 2017, 3:30 p.m.... construct. This strategy can be used to build vectors with diverse functions, and thus, is used in many...construction of plasmids with many components. The creators of the MXS-chaining method have used the technique...Shetty RP, Endy D, Knight TF. Engineering BioBrick vectors from BioBrick parts. Journal of biological engineering... -
Uncovering Ancestral Bias in CRISPR Research
TypeBlog PostPublishedNov. 29, 2022, 2:15 p.m....Biomedical research has a long and well-documented history of racial bias, which still impacts and influences...Further complicating the issue is the long and ugly history of reporting false essentialist links between biology...actively look for and consider bias as a confounding factor when interpreting findings, particularly when researching... -
Plasmids 101: Colony PCR
TypeBlog PostPublishedMay 12, 2016, 2:30 p.m....colonies. The best way to do this is by using your vector with and without an insert to verify that the primers...cultures of your positive clones. If you want to store your clones longer term, just streak the colonies...PCR primers work in the first place: the backbone vector with and without an insert. These controls are ... -
Plasmids 101: Protein tags
TypeBlog PostPublishedDec. 11, 2014, 4:26 p.m....number of chemical and peptide-based proteosome inhibitors, glutathione S-transferase (GST), which can be...purification to get your protein as pure as possible? Vectors that offer different combinations of tags are readily...Protein (GFP) Fluorescent Protein Guide Expression Vectors from the Berkeley QB3 MacroLab Eric thanks his... -
Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More
TypeBlog PostPublishedDec. 2, 2014, 7:23 p.m....applications? Fujii: We generated a series of retroviral vectors expressing tagged dCas9 and guide RNA. The retroviral...joined Addgene's Advisory Board, as well as the Editorial Board of Scientific Reports and PLoS One. Additionally...especially for young researchers. It was a really nice incubator for me. In Japan, of course, it is optimized for... -
Finding nucleic acids with SHERLOCK and DETECTR
TypeBlog PostPublishedApril 16, 2020, 1 p.m....time-consuming, and complex to use outside of standard laboratories. In the case of the COVID-19 pandemic, qPCR ...each enzyme. In addition, Csm6, a CRISPR type-III effector nuclease, can be used in conjunction with Cas13...single-component programmable RNA-guided RNA-targeting CRISPR effector. Science 353:aaf5573 . https://doi.org/10.1126... -
Antibodies 101: Immunoprecipitation
TypeBlog PostPublishedDec. 7, 2021, 2:15 p.m....Western blot. When choosing beads, there are several factors to consider. Agarose beads are sponge-like and ...more stringent buffers may be needed. Since many factors, including salt concentration, ionic strength and...prevent this, include protease and phosphatase inhibitors in the lysis buffer, keep samples on ice, and... -
CRISPR 101: Targeting RNA with Cas13a (C2c2)
TypeBlog PostPublishedSept. 21, 2017, 2:07 p.m.... (LshCas13a) to begin to characterize this new effector (Abudayyeh et al., 2016). In the endogenous CRISPR...fluorophore. The fluorescent signal can be used as an indicator to determine whether the target sequence is present...single-component programmable RNA-guided RNA-targeting CRISPR effector." Science 353(6299) (2016):aaf5573. PubMed PMID... -
Fluorescent Proteins 101: Aptamer Fluorophores
TypeBlog PostPublishedApril 11, 2017, 2:30 p.m....on FRET techniques. By combining standard donor/acceptor FRET know-how with aptamers, aptabeacons (or aptatags...fluorophore protein. Years later, Samie Jaffrey’s laboratory took advantage of their deep knowledge of the...a higher melting temperature than its Spinach ancestors, and has a lower magnesium dependence. Though ... -
Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design
TypeBlog PostPublishedSept. 13, 2018, 1:55 p.m.... delivered as a pooled library via a lentiviral vector to cell lines that either stably expressed Cas9... screen did fail to identify some known ricin-regulators, but most of these genes are also essential for...hypersensitivity, or are in an enhancer, transcription factor binding site, or a coding region. Targeting these... -
Tips for Writing a Good Cover Letter
TypeBlog PostPublishedMay 21, 2024, 1:30 p.m....logistical, like briefly explaining a gap in your job history or sharing relocation plans. They can be skills-based...in the possibility of joining your team as a postdoctoral researcher. Dear Hiring Manager, I am writing...your cover letter, and if possible, send it to a mentor or colleague for a second look and helpful feedback... -
Plasmids 101: Codon usage bias
TypeBlog PostPublishedSept. 27, 2018, 1:09 p.m....- e.g. codon choice may interrupt transcription factor binding sites) As you might imagine, it’s not ...not easy for humans to balance all of these factors on their own. Luckily, many researchers have created...Kimberly A., et al. "Selective charging of tRNA isoacceptors induced by amino‐acid starvation." EMBO reports... -
Using Video to Share Your Science: We Share Science
TypeBlog PostPublishedNov. 3, 2016, 2:30 p.m....research scientists, faculty, students, private sector researchers, or pretty much anyone applying the...studies on reducing antibiotic prescriptions for respiratory tract infections. The video abstracts provide...is not hard either. Here are some resources: ImpactStory, a website that helps researchers go beyond citation... -
Five Popular Model Organisms
TypeBlog PostPublishedApril 11, 2019, 12:39 p.m.... al., 2009). By injecting mice with retroviral vectors that harbored different mutations found in leukemia...Gerald Rubin’s lab who created a set of modular vectors that can be used to optimally control gene expression...discovery of RNAi in the worm actually rewarded depositors Andrew Fire and Craig Mello a Nobel Prize in ... -
Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior
TypeBlog PostPublishedMarch 5, 2019, 1:08 p.m....either by electroporation of plasmids, or using AAV vectors. The Imai lab also developed a version of Tetbow...Livet, Jean, et al. "Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous...Markus Grompe. "Clonal tracing of Sox9+ liver progenitors in mouse oval cell injury." Hepatology 60.1 (... -
Plasmids 101: Expressing shRNAs for RNA Interference
TypeBlog PostPublishedJune 17, 2025, 1:15 p.m....CRISPR, RNAi methods are often quicker and the repositories of validated small RNAs are vast and difficult...processing Following delivery into cells using viral vectors or plasmids, shRNAs hijack the endogenous miRNA...cloning strategies for constructing shRNA expression vectors. BMC Biotechnology, 6(1). https://doi.org/10.1186...