We narrowed to 393 results for: ara-2
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TypeBlog PostPublishedOct. 29, 2014, 5:26 p.m....Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...(old board games and movies can provide great character costumes). All of this costume planning is for...
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Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog PostPublishedSept. 8, 2016, 2:30 p.m....expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...cell 58.4 (2015): 644-659. PubMed PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve...: 26575238. 5. Crivat, Georgeta, and Justin W. Taraska. "Imaging proteins inside cells with fluorescent... -
Supporting Reproducibility with a Connected ELN
TypeBlog PostPublishedJan. 17, 2019, 1:38 p.m....connectivity to other data sources used in research; (2) connectivity to other tools used in research; and...by making it easy to get data out of the ELN; and (2) facilitate inter-operability with other research ... delve deeper and look specifically at the characteristics and capabilities an ELN needs to facilitate... -
Antibodies 101: The Basics of Western Blotting
TypeBlog PostPublishedMarch 8, 2021, 2:41 p.m....blotting In a western, proteins are: (1) separated by size, (2) transferred to a membrane, and (3) detected...system-specific sandwiches. Figure 2: The western blot dry apparatus includes a stack of filter paper, ...range to be separated on a single gel. Immobilizing proteins on a membrane Once separated, the proteins... process. The sample, a mixture of protein is separated in a gel, transferred to a membrane, stained with...tertiary structures remain intact and the protein is separated through a matrix by charge. In a denaturing western...protein is denatured to its primary structure and separated by size with smaller molecules moving more quickly... we will be focusing on denaturing westerns. Separating a protein mix by size Sodium dodecyl sulfate-... -
Antibodies 101: Beyond Surface Labeling
TypeBlog PostPublishedJune 27, 2024, 1:15 p.m....would label everything but the CD8+ T cells (Figure 2). Even better, we can use the same fluorophore for...gating on the negative population. Figure 2: Dump gating labels all unwanted cell lineages with...conjugated to a biotin tag but not a fluorophore. In a separate step, cells are incubated with a fluorophore-conjugated...the intranuclear ones. You do not need to use a separate fix/perm cytosolic procedure in that case. ...cell population. To detect non-surface markers, a separate intracellular labeling protocol is necessary, ... -
New Optogenetic Tools for Cytoskeleton and Membrane Control
TypeBlog PostPublishedSept. 28, 2023, 1:15 p.m....to a specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID ...cells will be ready for the spotlight. Figure 2: Opto-RhoGEFs to control Rho GTPase activity. A) Schematic...cytoskeleton. They provide cells with physical structure, separate chromosomes during mitosis, support transport ... called a katana. Opto-katanin consists of two separate blue light-sensitive systems, VVDfast to control...your protein of interest and express the other separately, then use blue light to generate CRY2-CIB1 oligomers... -
Six Spooky Science Stories and Halloween at Addgene
TypeBlog PostPublishedOct. 31, 2019, 1:06 p.m....melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed PMID: 15026169. Soo, Rochelle...algae host for reproduction. Snail parasite Leucochloridium is a parasitic worm that infects a snail’s eyestalks...Brusca and Gilligan, 1983). Grasshopper parasite The parasitic worm Spinochordodes tellinii‘s larvae makes...ve also carved pumpkins (yes, Blugene made an appearance there too) and shared fascinatingly creepy science...Tongue-eating louse Cymothoa exigua is a marine parasite that enters fish through its gills. It attaches...replacement in a marine fish (Lutjanus guttatus) by a parasitic isopod (Crustacea: Isopoda)." Copeia 1983.3 (1983... -
Mycoplasma Contamination: Where Does It Come From and How to Prevent It
TypeBlog PostPublishedOct. 8, 2020, 1:15 p.m....environmental contaminants. Figure 2: An Addgenie works in the tissue culture hood while wearing...doi.org/10.1290/1071-2690(2002)038<0086:caeomi>2.0.co;2 Uphoff CC, Drexler HG (2011) Detecting Mycoplasma ...of the causes of contamination. If possible, a separate lab should be made available for cell culturing...keeping them in the pockets of your lab coats. Have separate footwear for the lab to avoid bringing in environmental...wide range of mycoplasma. These methods cannot guarantee complete removal of mycoplasma but, can get rid...made permanent damage to the cell culture it is irreparable. Some cells may die due to longer incubation ...Microbiol .44: 42–49 Uphoff Cc, Drexler Hg (2002) Comparative Antibiotic Eradication of Mycoplasma Infections... -
Multiple Plasmids at a Low Price: Kits from Addgene
TypeBlog PostPublishedSept. 14, 2016, 2:30 p.m....wheat germ, HeLa, and Leishmania (Gagoski, et al 2015[2]). The proteins encoded in this set range in size ...to generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013. These terminators can...size from 4 to 156 kDa, enabling the user to characterise the efficiency of their cell-free system in correlation... -
Inntags: Innovative Protein Epitope Tagging
TypeBlog PostPublishedNov. 10, 2015, 3:30 p.m....tagging." Nature methods (2015). PubMed PMID:26322837. 2. Stadler, Charlotte, et al. "Immunofluorescence and...the fusions had some negative effect on these parameters. IT5 and IT6 displayed good binding efficiency... -
pSiM24: Simplifying Plant Genetic Engineering
TypeBlog PostPublishedSept. 29, 2015, 2:30 p.m....Agrobacterium infection. At approximately 7 kb, pSiM24 is 2 and 6 kb smaller than commercially available pCAMBIA...duplicated enhancer domains and characterized in studies of tobacco, Arabidopsis, and corn. For both β-glucuronidase...plant infection, as well as plasmid yield during preparation in E. coli. Addgene depositor Indu Maiti has ...functions in multiple species, including tobacco and Arabidopsis. Improving Agrobacterium binary vectors Vectors...efficiently inherited in both transgenic tobacco and Arabidopsis. Finally, Sahoo et al. demonstrate that this ... -
What's the Best Way to Elute and Store Your Plasmid DNA?
TypeBlog PostPublishedJune 4, 2020, 1:22 p.m....elution choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose ..., destabilizing the molecule and favoring the separation of the two helixes. Low pH instead results in... buffer for preserving the stability of your preparation for a long time. Tris buffer controls the pH,... if the priority is to directly use your DNA preparation in any downstream application involving enzymatic...buffer is that it allows better control of the pH preparation stabilizing the plasmid, and still allowing most...at -20°C or even at -80°C, in this case your preparation can be eluted in water or in your buffer of preference... -
CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?
TypeBlog PostPublishedOct. 13, 2022, 1:45 p.m....sequence is unique compared to the rest of the genome. 2) The target is present immediately upstream of a Protospacer...utilized allowing for PAM flexibility. Fig. 2: Prime editor engaging target DNA. The reverse transcriptase...Walton et al., 2020). Further, this group also characterized a second Cas9 variant, SpRY, which cuts at NRN... -
pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs
TypeBlog PostPublishedDec. 14, 2017, 2:08 p.m....blood are reprogrammed immediately after collection [2]. PBMCs display unique epigenetic signatures that ...PMID: 27882265. PubMed Central PMCID: PMC5118044. 2. Kim, Y., et al., The Generation of Human Induced ...vectors encoding reprogramming factors behave as separate extrachromosomal elements in the nucleus of the... -
Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility
TypeBlog PostPublishedMay 18, 2018, 7:32 p.m....Almo, Institute for Protein Innovation 1:18:45 - 2:01:18 (end) - Panel Alex Tucker (Ginkgo Bioworks)...their curation of citable, linkable figures. Cause #2: Hidden “negative” data We all want to be able to ... and here I summarize what I learned both in preparation for my talk and at the minisymposium. You can...address this issue directly by creating well characterised antibodies against all human extracellular proteins...stories and read papers wherein a researcher tries 5 separate experiments at once to get confirmatory data for... -
CRISPR 101: Validating Your Genome Edit
TypeBlog PostPublishedNov. 3, 2022, 12:15 p.m....faster when using a dual guide system. Fig. 2: Size screening for large deletion events. ... Biol 17: https://doi.org/10.1186/s13059-016-1012-2 Find more CRISPR Resources at Addgene: Read our CRISPRGuide...visualized by running your PCR on a gel. It’s still paramount that you sequence clones to verify that the deletions... -
Quick, Versatile Plant Transgenesis with GreenGate Plasmids
TypeBlog PostPublishedJan. 23, 2014, 3:23 p.m....Transgenesis." PLoS ONE. 8(12): e83043 (20 December 2013). 2. Carola Engler et al. "A One Pot, One Step, Precision...Lohmann said they’ve only tested GreenGate in Arabidopsis, it should in principle work in any plant species... -
Let There Be LITE Plasmids
TypeBlog PostPublishedNov. 21, 2013, 3:56 p.m....First, the light-responsive molecules of cryptochrome 2 are the chromophores flavin-adenine dinucleotide and...ingredients - CRY2 and CIB1 – which they borrowed from Arabidopsis thaliana. The results are light-inducible transcriptional... -
Custom CRISPR Screens & the Green Listed Software
TypeBlog PostPublishedJuly 11, 2017, 2:30 p.m....six different research constellations: Doench/Root (2), Zhang (3), Wu (4), Yusa (5), Wang/Lander/Sabatini...Bioinformatics 33.7 (2016): 1099-1100. PubMed PMID: 28414855. 2. Doench, John G., et al. "Optimized sgRNA design to...single cell level), are separated by, for example, cell sorting. The separated cells are subsequently ...which gRNAs are differentially expressed in the separate groups and, as a consequence, which genes are ...the protein following stimulation would then be separated by sorting, and subsequently sequenced to identify...suggested gRNA sequence including adapters. Output_UserInputParams: Contains all the information that you ...al. "Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach." Nature methods... -
Finding Your Science Policy Path
TypeBlog PostPublishedSept. 22, 2020, 1:15 p.m....teaching and mentoring others (see more in Figure 2). Starting early demonstrates not only your interest...application process so you can learn more! Figure 2: The many ways to prepare for a science policy career...the work you do to creatively connect seemingly disparate individuals and ideas toward solving one problem...evidence-based university policy to combat sexual harassment in academia or a large company who decides how...