We narrowed to 512 results for: ato
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TypeBlog PostPublishedOct. 18, 2018, 12:37 p.m.... Javier García-Nafría is an investigator scientist at the MRC Laboratory of Molecular Biology. He is ... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential...with successful recombination reported in all laboratory E. coli strains tested so far. The use of this...low cost and can be immediately adopted by any laboratory. All procedures are performed using the same...the homologous region (calculated using Oligo Calculator). Insertions - To insert short sequences of ... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Jake Watson is a postdoctoral...postdoctoral researcher in neurophysiology at the MRC Laboratory of Molecular Biology. He is currently investigating...
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6 Tips for Analyzing and Troubleshooting Sanger Sequencing Results
TypeBlog PostPublishedJune 26, 2014, 2:23 p.m....to always look closely at the trace file (or chromatogram) of the sequencing results you get back from...facility. When it comes to DNA sequencing the chromatogram is your visual control. And, like all controls...controls, missing out is a big mistake. Chromatograms come to the rescue for analyzing and troubleshooting...But look closer, the overlapping peaks in the chromatogram suggest the results are not as certain as the... of the following programs to view your .ab1 chromatogram file 4Peaks (Mac) SnapGene Viewer (Mac/PC) ...peak, don’t be shy. Feel free to edit it. Most chromatogram viewing programs (even the free ones) allow ...allowing us to share it with our readers! The chromatograms in this article were created with SnapGene. ... -
Hot Plasmids - August 2020
TypeBlog PostPublishedAug. 18, 2020, 1:15 p.m....mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato, mDlx-GCaMP6f, hDlx-GiDREADD-dTomato and mDlx-ChR2-mCherry. Find... a cell. The Goedhart lab, Gadella lab, and collaborators have an ongoing effort to improve genetically...by one-step immobilized metal ion adsorption chromatography (IMAC) using Ni-NTA magnetic beads. Workflow...multiplexed gRNAs and of dCas9 fusions. The dCas-KRAB activator and the dCas-VPR repressor are introduced into...an EGFP reporter. Expression of the dCas9-KRAB activator or dCas9-VPR repressor in hPSCs is also driven...expression. This includes a CRISPRi-based synthetic oscillator, bistable network, stripe pattern-forming incoherent...calcium sensors (jYCaMP1), variants of the calcium indicator jGCaMP7, that are optimized for excitation wavelengths... -
Plasmids 101: The Promoter Region – Let's Go!
TypeBlog PostPublishedApril 3, 2014, 8:05 p.m....controlled by still other regulatory factors: enhancers, boundary elements, insulators, and silencers; however...tetracycline operators. Transcription can be turned on or off depending on what tet transactivator is used....the promoter (RNA polymerase binding site) and operators (response elements). Promoters are about 100 to...expression Promoter from T7 bacteriophage plus lac operators Negligible basal expression when not induced. ...RNA polymerase, which is also controlled by lac operator. Can be induced by IPTG. Commonly found in pET...pET vectors. Very tightly regulated by the lac operators. Good for modulating gene expression through varied... -
Tetbow: Bright Multicolor Labeling for Neuronal Tracing
TypeBlog PostPublishedJan. 24, 2019, 2:24 p.m.... introduced, stochasticity will result in a combinatorial expression of these three genes with different...inherently low. This is because the stochastic and combinatorial expression of fluorescent proteins is only possible...original method, we used the Tetracycline trans-activator system to boost the expression levels, and we ...we named the system Tetbow (Tetracycline trans-activator Brainbow) (Sakaguchi et al., 2018) (Figure 1)....sensory systems focusing on the functional and anatomical maturation of the olfactory system. Takeshi...University. His lab has been studying functional and anatomical aspects of sensory circuit development using ...Livet, Jean, et al. "Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous... -
Cultivating Community Science at BosLab
TypeBlog PostPublishedNov. 2, 2017, 1:35 p.m....fully-equipped laboratory for citizen scientists. In addition to maintaining a laboratory to support both... of independent Do-it-Yourself (DIY) biology laboratories working towards enabling everyone to participate... set on top of filing cabinets, a rickety old incubator, a few pipettes, and some home-made equipment....Anyone can become a member of BosLab and previous laboratory experience is not required. Our members are skilled...have non-traditional scientists working in our laboratory! Don’t worry though, we make sure that everyone...a co-organizer at BosLab, a community biotech laboratory in Somerville MA. Follow her on twitter at @ZebraElement... -
Mycoplasma Contamination: Where Does It Come From and How to Prevent It
TypeBlog PostPublishedOct. 8, 2020, 1:15 p.m....include the laboratory personnel, the serum, the cell culture media, water baths, incubators, etc. Contamination... in a single incubator since it increases the risk of cross-contamination. CO2 incubators should be cleaned...Contamination! Tissue culture media often have indicators included in the media that turns yellow from ...proliferating shortly. Once they are present in the laboratory environment, they are difficult to eradicate....tighten the flask cap before placing them in the incubators. This will also solve the problem of contamination...airflow and the lab facility periodically. The CO2 incubators, used for maintaining cell lines, must not be... -
Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards
TypeBlog PostPublishedAug. 5, 2021, 12:15 p.m....tag neurons activated by opioid withdrawal with tdTomato and assessed cFos expression. She will explore...periaqueductal gray (PAG), with red neurons expressing tdTomato and green neurons expressing cFos. Image from ...painful stimuli are processed by a network of excitatory and inhibitory neurons whose function and connectivity...where she engineers genetically encoded voltage indicators. Tian developed a video-based high-throughput...archaerhodopsin‑derived genetically encoded voltage indicator which had improved performance in live mouse brains... Currently, she is using the improved voltage indicator coming from the screen to understand mouse cortical...Tian. Figure 5: Citrine-tagged voltage indicator expressed in mouse cortex. Image from He Tian.... -
Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics
TypeBlog PostPublishedMarch 3, 2020, 2:15 p.m....voltage indicator Article contributed by Shreya Vedantam Genetically encoded voltage indicators (GEVIs)...fluorescent optics. Existing red fluorescent voltage indicators, however, fall short compared to GFP-based sensors... sensors and blue-light-activated optogenetic actuators. To overcome this overlap, Yiyang Gong’s lab ...linker between them. The Ace-mScarlet voltage indicator works like many FRET-opsin GEVIs. With an increase...channelrhodopsins. Find the Ace-mScarlet voltage indicator plasmids at Addgene! Beck et al., Sci Report 2019...cyanobacteria. This kit contains 96 parts including terminators and promoters, CRISPRi parts, selectable markers... -
Cancer and the Immune System: Deciphering the Relationship
TypeBlog PostPublishedMarch 14, 2017, 2:30 p.m....and helper T (Th) cells. These cells produce inflammatory cytokines - interleukin (IL)-12 and interferon... eliminate early stage cancer cells, chronic inflammatory responses can augment tumor growth. Indeed, ... to precede cancer development, for example, inflammatory bowel disease is associated with the onset of... 1 (CSF-1). Additionally, tumor cells recruit regulatory T cells (T-regs) into their microenvironment ...TGFβ is also known to change CD4+ Th cells into regulatory cells to suppress function. Creating Tolerance...cell-surface proteome to inhibit expression of co-stimulatory molecules needed for T cell effector function...identities based on their level of progression, anatomical location, characteristic (solid localized tumors... -
Viral Vectors 101: Calcium Sensors
TypeBlog PostPublishedAug. 10, 2023, 1 p.m....crosslinking agent, a signaling ion, and an allosteric modulator. Because of its importance, scientists have developed... a calcium sensor Genetically encoded calcium indicators (GECIs) are fluorescent proteins linked to calcium...in living cells and organisms. Unlike chemical indicators, these DNA constructs can be delivered to intact...:509-18. Inoue M. Genetically encoded calcium indicators to probe complex brain circuit dynamics in vivo...Kanemaru K, Iino M. Genetically encoded fluorescent indicators for organellar calcium imaging. Biophysical journal...neuronal activity with genetically encoded calcium indicators. Cold Spring Harbor Protocols. 2012 Jun 1;2012...signaling using genetically targetable fluorescent indicators. Nature protocols. 2006 Aug;1(3):1057-65. Resources... -
Tips to Make the Most of a Scientific Conference
TypeBlog PostPublishedMay 23, 2019, 12:26 p.m....following: the elevator pitch, talks, the poster session, networking, and social media. The elevator pitch Before... Before the conference Practice your “elevator pitch,” the 30 - 60 second blurb you will use to introduce...work you are familiar with. You can tweak your elevator pitch slightly if you know a bit more about the...or talk. After the conference After using your elevator pitch several times at the conference, you might...might notice some trends. Were there parts of your elevator pitch where the other person seemed confused, ...questions. If you or your lab identifies a potential collaborator from the meeting, reach out to them, pointing... -
3D Printing Meets CRISPR Cas9
TypeBlog PostPublishedOct. 17, 2017, 12:55 p.m....development programs for bioscience educators. Using these hands-on tools, educators introduce their students to...a backbone structure of a protein, based on its atomic coordinates. These wire models were useful in that...necessary to download a pdb file of the structure’s atomic coordinates, and “open” these structures with molecular...dangerous. Subscribe to CRISPR Blog Posts How do educators and students use molecular models? We created...something tangible. Under the guidance of a trained educator, these instances of focused attention become magical... their labs. Soon after we began working with educators in our Center for BioMolecular Modeling, we spun... -
CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]
TypeBlog PostPublishedFeb. 18, 2015, 3:09 p.m....The first is the study of non-coding regulatory elements. Regulatory DNA function is typically inferred ...bloggers! Daniel Bauer is a pediatric hematologist and principal investigator at Boston Children’s Hospital and...interested in exploring the function of non-coding regulatory elements in their native chromatin context. An...PhD student at Harvard Medical School in the laboratories of Stuart Orkin and Daniel Bauer. He received... -
Technologies Enabled by NanoLuc® Luciferase
TypeBlog PostPublishedFeb. 8, 2018, 12:17 p.m....employing optogenetic regulators to initiate the event being monitored. Optogenetic regulators respond to specific...initiation wavelengths, excitation of the optogenetic regulator leads to spurious activation of the FRET sensor...interested in using photoactivatable, optogenetic accuators to depolarize or hyperpolarize membranes. Traditional...wavelengths needed would also activate the optogenetic accuators. The researchers took a FRET reporter and adapted...biosensors that can even be paired with optogenetic regulators. Many of these tools are available from Addgene...2017) Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of bioimaging... -
The Crispr Babies Saga Shows the Need for Action, Not More Delays
TypeBlog PostPublishedDec. 20, 2018, 1:15 p.m....upon a moratorium until another meeting, three branches – a private funder, government regulators, and ...there was a generally agreed-upon international moratorium on utilizing CRISPR/Cas9 in humans. This topic...mtDNA transfer options being investigated; A UK regulatory body tasked two separate non-governmental groups...technology to research centers engaged in long-term, mandatory follow-ups and observational studies of the technique...stop punting decisions on scientific, ethical, regulatory, and general public consensus to the next meeting... -
Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools
TypeBlog PostPublishedDec. 10, 2019, 1:58 p.m....lipoproteins, shuttles lipoproteins through the circulatory systems, but can also lead to the formation of...as both a powerful tool for generating complex regulatory systems and further the study of natural σ54-... PMID: 31451697. Genetically encoded voltage indicators for use in worms Article contributed by Brook...Brook Pyhtila Genetically encoded voltage indicators (GEVIs) change fluorescence in real time when neurons... excitation light intensities have made these indicators difficult to work with. The Gottschalk lab has...microbial rhodopsin GEVIs in excitable cells of the nematode Caenorhabditis elegans. They have found several... -
Plasmids 101: Modular Cloning Applications and Kits
TypeBlog PostPublishedMay 14, 2024, 1:15 p.m....site-specific recombinases, regulatory elements, developmental regulators, and lots of viral components...test many different combinations of promoters, terminators, solubility and purification tags, connectors... examine the effects of different promoters, terminators, enhancers, and cross-cassette interactions in...Standard and New E. Coli Part Library Enable Rapid Combinatorial Design for Synthetic and Traditional Biology... Signaling in CAR T Cells. Cold Spring Harbor Laboratory, 17 Feb. 2024. Crossref, https://doi.org/10.1101...Emerging CRISPR Technologies. Cold Spring Harbor Laboratory, 27 Oct. 2023. Crossref, https://doi.org/10.1016... -
The Breast Cancer Microenvironment: A Tumor’s Backstage Team
TypeBlog PostPublishedJan. 31, 2019, 1:32 p.m....environmental modulation of hepatocellular cancer cell growth." Hepatology 54.4 (2011): 1237-1248. PubMed... was contributed by Bárbara Pinho, a science communicator at the Portuguese science museum "Fábrica Centro...myofibroblasts, neuroendocrine, adipose, immune, inflammatory, angiogenic vascular, endothelial, and stromal...seems to attack us by releasing mitogenic growth mediators and improving cancer cells proliferation. How... Pinho! Bárbara Pinho is currently a science communicator at the Portuguese science museum "Fábrica Centro... -
Hot Plasmids - October 2020
TypeBlog PostPublishedOct. 13, 2020, 1:15 p.m....you'll find in this post: Tools for labeling excitatory inhibitory synaptic proteins Fluorescent protein...corner New from the viral service Labeling excitatory and inhibitory synaptic proteins with FingRs ...developed AAV vectors to selectively visualize excitatory and inhibitory synapses in the brain. ...scaffolding proteins PSD95 and gephyrin to label excitatory and inhibitory synapses, respectively. Once expressed... fluorescent proteins targeting inhibitory or excitatory synapses or that are Cre-inducible, researchers... cycle in vivo, particularly if the cells are migratory or in deep tissues. To resolve this issue, Shangqin...