We narrowed to 208 results for: PHI-1
-
TypeBlog Post...optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference... plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan, A., ... used arg1 to in vivo image tumour-homing S. typhimurium strain ELH1301. Bourdaeu et al see this as just...
-
Antibodies 101: Single Chain Fragment Variables (scFvs)
TypeBlog Post...the other (Sandomenico et al., 2020). Figure 1: Comparison between the IgG antibody and scFv. ...intensity of a tagged protein. SunTag has two components: 1) a protein of interest that’s tagged with 10-24 copies...Application. Clinical and Developmental Immunology 2012:1–15 . https://doi.org/10.1155/2012/980250 Bates A,...without disulfide bonds, made by molecular evolution 1 1Edited by I. A. Wilson. Journal of Molecular Biology...made up of glycine and serine with dispersed hydrophilic residues for increased solubility (Monnier et... -
Six Spooky Science Stories and Halloween at Addgene
TypeBlog Post...biosynthesis in Drosophila melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed...stories. Six spooky science stories Zombie ants Ophiocordyceps unilateralis is a fungus that infects ants ... a set of cytochrome P450 genes discovered in Drosophila melanogaster (Gilbert, 2004). Mutations in these...blood out. Once the blood is depleted, the tongue atrophies and falls off. The louse remains becoming the ... -
Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins
TypeBlog Post...References 1. Peters, Nathan C., et al. "In vivo imaging reveals an essential role for neutrophils in leishmaniasis...do researchers track fluorescently labeled cells? 1. Epifluorescence microscopy This technique uses a ... The video above depicts the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph...infection with S. aureus (red). You can then see neutrophils swarming around S. aureus and eating the bacteria...two-photon microscopy include: How do T cells and/or Neutrophils travel to the sites of infection or damage (Peters...Good two-photon probes when using a standard Ti:Sapphire Laser include: Blue-green fluorescent proteins...this requires other lasers than the standard Ti:Sapphire laser. It is also important to finely tune the... -
CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments
TypeBlog Post...This gives each guide a score ranging between 0 and 1. The cutting frequency determination (CFD score) is...converted to a weight (which ranges between 0 and 1) so that they can be used in the rank equation above...converted to a weight (which ranges between 0 and 1) (shown in the graph below) so that it can be used...HDR. Optimal mutation in-depth One the most sophisticated features of CrispyCrunch is how it mutates guide... -
Which Fluorescent Protein Should I Use?
TypeBlog Post...biological questions. The two most common uses are: 1) to test the expression level in a specific system...brightness is compared to that of EGFP which is set as 1. Some proteins are very dim (e.g. TagRFP657, which...Photostability can be as short as 100ms (EBFP) or as long as 1 hour (mAmetrine1.2). However, for most FPs it is a...emission is green or red light. For example, T-Sapphire, LSSmOrange, and LSSmKate. Fluorescent Sensors... -
Early Career Researcher Toolbox: Free Online Molecular Biology Tools
TypeBlog Post...more tools for plasmid mapping and design (Figure 1). Benchling: While you might think of Benchling as...Davis from the University of Utah. Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid...patterns, post-translational modifications and polymorphisms Codon optimization tool: To access this tool... -
Hot Plasmids and Viral Preps - July 2021
TypeBlog Post...organoid model (Sun, et al. 2021). Figure 1: Example schematic from the Organoid EasyTag system...without altering that of the internal control (Fig 1). Because this method relies on the fluorescence ratio...and can be used for genome editing in moderate thermophiles. These evolved SpCas9 variants have improved... -
A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light
TypeBlog Post... that dimerizes upon exposure to blue light (Fig. 1). Another blue light-inducible dimerization domain...expression needed for most applications. Figure 1: With blue light, the VVD domains of the BLADE dimerize...Mustafa Khammash's lab at ETH Zürich. “Instead of a graphical abstract, it’s a musical abstract,” says Di Ventura... -
Summer SciComm: Science Storytelling
TypeBlog Post.... In literary terms, conflict can be defined as (1) the struggle between two opposing forces or (2) the...journey even when they know the ending. Fig. 1: We all know that the superheroes win in the end. ...other affected people, man/animal vs nature, anthropomorphized animals, or even seemingly unrelated stories... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog Post...tethered to bring them into close proximity (Figure 1). Formation of the complete FP results in a fluorescent...detected by fluorescence microscopy. Figure 1: How BiFC works. A split FP (e.g., yellow fluorescent... protein interactions in living cells. Nat Protoc 1:1278–1286. https://doi.org/10.1038/nprot.2006.201 ...analyzing protein–protein interactions in live Drosophila. eLife 7. https://doi.org/10.7554/elife.38853... -
Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs
TypeBlog Post...experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable researchers ... disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs that encode N-glycosylation... sarcoma-associated herpesvirus. References 1. Snapp, Erik. "Design and use of fluorescent fusion...genes: deletion mapping functional domains of Drosophila hsp 70." The EMBO journal 3.13 (1984): 3087. ... -
Your Top Requested Plasmid in 2015!
TypeBlog Post...another great year of sharing! References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout...Central PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang. "Improved vectors and genome-wide... -
History of CRISPR Cas - A tale of survival and evolution
TypeBlog Post...sequences were initially discovered in E. coli in 1987 (1), the concept that these clustered repeat sequences...neutralize CRISPR/Cas. Earlier this year, expression of 1 of 5 anti-CRISPR genes were found to inactivate the...light until 2007. Initial experiments exposed S. thermophilus to predatory phages to test if the exogenous... -
Plasmid Cloning by PCR
TypeBlog Post...has error rates that range from roughly 1 per 500bp to roughly 1 per 10 million bp depending on the polymerase...analyzing the gene’s function. The process is shown graphically in the following figure, in which we add EcoRI... -
Tag Your Favorite Yeast Genes with Ease
TypeBlog Post...vectors and include a Kan, SpHIS5, or CaURA3 selection marker. Lee et al.(1) assessed many of these fluorescent... Shah NG, McKenzie A 3rd, Steever AB, Wach A, Philippsen P, Pringle JR. Yeast. 1998 Jul;14(10):943-51....3rd, Demarini DJ, Shah NG, Wach A, Brachat A, Philippsen P, Pringle JR. Yeast. 1998 Jul;14(10):953-61.... -
Hot Plasmids: Fall 2024
TypeBlog Post...construct for crystallographic and biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https...and is recommended for use in western blots (Figure 1). If you prefer to produce your own antibody or want...are also available from Addgene! Figure 1: Experimental validation for Anti-V5 [IPI-SV5-Pk1]... -
CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?
TypeBlog Post...guide RNA (sgRNA), provided it meets two conditions: 1) The sequence is unique compared to the rest of the...bystander bases” if you choose this system. Fig. 1: Cytidine deamination takes place on the free strand...temperature-controlled genome editing. Nat. Commun. 2017 81 8, 1–9 (2017).Walton, R. T., Christie, K. A., Whittaker...(but not limited to) bacteria, yeast, plants, drosophila, worms and mammals (Browse plasmids by model ... -
Important Considerations in Optogenetics Behavioral Experiments
TypeBlog Post...animal will repeatedly nose-poke at the active hole [1, 2]. Place preference/place aversion also models... can be explained primarily in two different ways 1) your hypothesis is wrong or 2) there is some problem...follow him on Twitter @derekpsimonphd. References 1. Stuber, Garret D., et al. "Excitatory transmission...technology is improving and may one day reach the sophistication of mouse transgenics. Everything that has been... -
Site Directed Mutagenesis by PCR
TypeBlog Post...sequence on either side of the desired mutation (usually 1-3 mismatched bases) is sufficient for your primers...researcher at Cornell University. References 1. Breslauer, Kenneth J., et al. "Predicting DNA duplex... PCR followed by Restriction Fragment Length Polymorphism (RFLP) analysis (right). Introduction of restriction...of that particular site with fragment length polymorphism (RFLP) analysis. In this process, if your mutation...