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...beginning Let’s take a quick flash back to 2004. “Shrek 2” was the most popular movie in theaters, the Razr ...amazing years in front of us. So in our last celebratory moment, we wanted to walk through some of our...Facebook and Gmail, were launched. And Melina Fan, a cell biology graduate student, was frustrated by the ...to receive the Massachusetts Nonprofit Network Excellence Award for our electronic MTA system. We also ...now our Sr. Director of Production) pie-fully celebrating Addgene's 10 year anniversary.   Expanding...

...method 2): Method 1 Method 2 $$T = {N*F*D\over V_T}$$ Where: T = Titer, TU/mL N = Number of cells transduced...Workflow Timeline Day 0: Seed 293T cells Day 1: Transduce cells Day 2 (am): Remove media, replace with ...,000 cells into 14 mL of DMEM complete. Mix well by pipetting or inverting. Aliquot 2 mL of cell suspension...fluorescent cells Equipment Class II, Type A2 Biological Safety Cabinet 0.5–10 µL single channel pipette 2–20 ...Lentivirus is generally considered biosafety level 2+. Please ensure that you are in compliance with your... 200–1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container...added to 150,000 cells resulted in 25% fluorescent cells, then the titer is: (150,000 cells x 0.25 fluorescent...

...localizations. Figure 2: Split-wrmScarlet labeling proteins with different subcellular locations. Image from...determine cell cycle length by examining the ratio of blue to red cells. Cells undergoing rapid cell cycles...engineered C. elegans cell lines to express wrmScarlet1-10 unattached to any other cellular protein. They used...’s difficult to observe the cell cycle in vivo, particularly if the cells are migratory or in deep tissues... BaF3 cells were transduced with H2B-FT virus and grown in varying concentrations of IL-3. Cells grown...proportion of blue fluorescence, indicating cell proliferation. BaF3 cells maintained in media without IL-3 show...H2B-FT consistently predicted the cell cycle speed across these different cell types. In one example, H2B-FT...

... already have an end product in mind, steps 1 and 2 will generally be straightforward. Likewise for step...Calcium phosphate Cells Transformed cell lines Transformed cell lines Transformed cell lines Cargo Nucleic...identify and enrich positive cells or a selection marker to generate stable cell lines. Transient or stable...and enrich positive cells. Likely contains a selection marker to generate stable cell lines. Packaging and...wide variety of mammalian cell lines. Useful for difficult to transfect cell types and can be used in ...protein are degraded within the cell.   Delivery methods for mammalian cell lines As mentioned above, the...components into your target cells. Delivery of these components into mammalian cell lines is quite broad and...

... tabular HTML formatted output. Surprisingly only 2 tools provide FASTA output. A few notable tools allow...Nature biotechnology(2014). PubMed PMID: 24880488. 2. Hsu, Patrick D., et al. "DNA targeting specificity...the research community. I’ve likened it to the celebration surrounding the piñata. From the perspective ...

... Cure for HIV/AIDS. Int J Mol Sci. 2023 Jan 13;24(2):1563. doi: 10.3390/ijms24021563 Resources on Addgene.org...Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients." Cell Stem Cell. 13, 653-658 (2013). ...patient’s own cells that have been modified ex vivo, previously edited healthy donor cells can be delivered...Genetic Disease in Mouse Via Use of CRISPR/Cas9." Cell Stem Cell. 13, 659-662 (2013). Gerald Schwank et al. ...on editing animal models of disease and cultured cells. For example, in 2013, Jinsong Li and colleagues...applied CRISPR to disease correction in adult stem cells isolated from two patients with cystic fibrosis ...delivery directly to the patient. Patient-derived cells can alternatively be harvested, edited ex vivo, ...

...protein from cells or tissue samples. Equipment Microcentrifuge 0.5–10 µL single channel pipette 2–20 µL single...transfer, block, incubate with primary antibody Day 2: Incubate with secondary antibody Video Watch this...Heat block Mini gel tank chamber Power supply iBlot 2 Gel Transfer Device Roller Spatula Platform shaker...running buffer Prestained protein ladder Ethanol iBlot 2 PVDF Mini Stack, Thermo Fisher IB24002 20X TBS Tween...immediately or store at -80 °C until ready to use. Section 2: Determine the total protein concentration and prepare...in deionized water. To prepare 20% ethanol, dilute 2 mL of ethanol into 8 mL of deionized water and mix...the transfer sandwich as follows: Unseal the iBlot 2 PVDF Mini transfer stack. Set the Top Stack to one...

...C-terminus of human codon-optimized  cryptochrome 2 (CRY2) from Arabidopsis thaliana. When exposed to ...illumination of neighboring cells evokes action potentials (APs) in patched cells expressing wild type CoChR...techniques for live-cell imaging of the dynamic changes in biochemical activity within the cell. Their Fluorescence...distribution of clusters without accumulation in subcellular compartments such as nuclear speckles. Furthermore...OptoSTIM1), caused two-fold faster changes in intracellular Ca2+ levels than the original version containing...CRY2clust should prove a useful tool to further probe cellular function. The CRY2clust Plasmid can be found here...28646204. soCoChR: A novel opsin that enables single-cell optogenetics Article contributed by Meghan Rego ...

...dimerization-dependent protein interactions. FEBS letters, 591 2, 415-424. PubMed PMID: 28032891. This is a different... expressed in cells via plasmids or by the creation of stable cell lines and these cells are then grown... complex. Not performed in a native cell context. For AP-MS, cells are lysed before the bait protein is...and interacting proteins in mammalian cells. The Journal of cell biology. PubMed PMID: 22412018. PubMed...against PPIs that aren't stable outside the native cell context. The Split-BioID method Enter Split-BioID...tagging of proteins in both BioID or Split-BioID, cells are lysed, protein extracted, and labeled protein...proteins that interact with the bait in a native cell contact. Other PPI methods may require protein interactions...

...divided by the off-target activity. Figure 2: Specificity is the measure of off-target activity...CRISPR-Cas12b for mammalian genome engineering." Cell discovery 4.1 (2018): 63. PubMed PMID: 30510770....