We narrowed to 845 results for: CAL;
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TypeBlog PostPublishedJune 23, 2015, 8:37 p.m....type; however, their usage has been limited by technical problems. For instance, it’s much more difficult...presence of the CPP may alter protein function or localization. Thus, the usefulness of CPPs must be evaluated...
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Mentoring for Scientists: I Have a Mentor, What Now?
TypeBlog PostPublishedMarch 6, 2014, 7:12 p.m.... Listen to our podcast interview with Harvard Medical School researcher Connie Cepko to learn about her...mentoring meetings a priority. Schedule meetings or calls regularly and don’t cancel, so that you drive progress... -
Extracurricular Activities for a Strong Science Career Path
TypeBlog PostPublishedJan. 21, 2014, 3:08 p.m....developed a web-based, career-planning platform called the Individual Development Plan (IDP) and uses ... Outreach activities (e.g. Volunteer at a local high school science class: Science Club for Girls... -
How Dry Ice Affects Viral Vector Sample pH & How to Avoid It
TypeBlog PostPublishedJune 20, 2017, 2:30 p.m....aqueous samples might be affected by dry ice, and specifically how dry ice can affect virus from Addgene. ...never stored on dry ice/exposed to CO2 (Fig. 5). Basically, the CO2 goes in and the CO2 goes out. Conclusions... -
Components of CRISPR/Cas9
TypeBlog PostPublishedFeb. 2, 2016, 5 p.m.... to modify the desired genomic locus. Some historical notes CRISPR arrays in bacterial genomes consist...arrays, they did not know their functions and simply called the repeated elements “direct repeats” and the ... -
Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2
TypeBlog PostPublishedMay 26, 2020, 1:15 p.m....end joining (NHEJ). In these reporters, editing typically results in a change in the fluorescent readout...stimulated homology-directed repair. Journal of Biological Chemistry 292:6148–6162 . https://doi.org/10.1074... -
Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors
TypeBlog PostPublishedMarch 17, 2020, 1:32 p.m.... specific DNA base changes and consist of a catalytically impaired Cas protein (dCas or Cas nickase) fused...previously uneditable sequences. Four DNA base swaps, so called point mutations that change C to T, G to A, A to... -
Hot Plasmids - February 2022
TypeBlog PostPublishedFeb. 22, 2022, 2:15 p.m....anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-GFP labeling...4xFLAG-mCherry-β-actin), and mitochondrial (mito-mCherry-1xFLAG) localization. Adapted from Liu et al. 2021. Anti-FLAG... -
Improved Plasmid Maps Powered by SnapGene
TypeBlog PostPublishedJune 29, 2017, 1:06 p.m....powered plasmid maps far easier to navigate. These practical and effective changes will give you the ability... and “Sequence” tabs. BLAST Use the Basic Local Alignment Search Tool (BLAST) via the NCBI website... -
How to Keep a Lab Notebook for Bioinformatic Analyses
TypeBlog PostPublishedJune 2, 2016, 2:30 p.m.... the following, which are later printed for my physical lab notebook, along with any figures generated...between each word can make it difficult to accurately call a file. Version control your commonly used scripts... -
Flow Cytometry Readouts: Yes, No, and Everything in Between
TypeBlog PostPublishedAug. 20, 2024, 1:15 p.m.... a downregulation of signal, though it’s hard to call from this plot. The red (bottom) peak overlaps with...Research Assistant in Arlene Sharpe’s lab at Harvard Medical School. More resources on the Addgene blog Antibodies... -
Viral Vectors 101: Gamma-Retroviral Packaging Systems
TypeBlog PostPublishedAug. 6, 2024, 1:15 p.m....transfer plasmid sequences into the host genome. Typically, it is the sequences between and including the...pseudotyping. Figure 1: Genome of a prototypical gamma-retrovirus. Frequently asked questions... -
Antibodies 101: Flow Cytometry Controls
TypeBlog PostPublishedJune 6, 2024, 1:50 p.m....sample. The isotype antibody will be virtually identical to the antibody for your marker of interest. It...research technician in Arlene Sharpe's lab at Harvard Medical School. More resources on the Addgene blog Introduction... -
Summer SciComm Series: Cognitive Load
TypeBlog PostPublishedJune 16, 2022, 1:15 p.m....intuitive, or logical to a biologist that are not at all obvious, intuitive, or logical to a layperson... -
CRISPR 101: Cas9 vs. The Other Cas(s)
TypeBlog PostPublishedDec. 8, 2022, 2:15 p.m....once it is engaged by its target, it will non-specifically cut ssDNA. This feature is shared by other Cas... potential as a viral therapeutic that could specifically degrade target viral RNA. For more information... -
Transferable Skills Guide: Cross-team Communication
TypeBlog PostPublishedNov. 21, 2019, 1:53 p.m....out the sources of miscommunication and b) find practical solutions. My PhD prepared me remarkably well...answer to these questions--which were real, not hypothetical--was clearly “no.” When there was room for misinterpretation... -
New Tool for Lineage Tracing: The ClonTracer Library
TypeBlog PostPublishedSept. 22, 2015, 2:30 p.m....library, propagated it, separated it into roughly identical replicate groups, and treated these groups with...would have different labels - i.e. they would stochastically arise from different cells throughout the treatment... -
Plasmids 101: Golden Gate Cloning
TypeBlog PostPublishedAug. 27, 2015, 2:30 p.m....Another strength of Golden Gate cloning is its scalability. Unique 4 base overhangs can be used to assemble...design of flanking overhangs. Although there are theoretically 256 distinct flanking sequences, sequences that... -
Career Coaching for Scientists: Why and Where Do I Find One?
TypeBlog PostPublishedJan. 29, 2015, 1:25 p.m.... will provide an initial consultation meeting or call for free. Take advantage of this to get the information...blogged for us) offers a free report on her website called “Secrets to Success in Graduate School”. She has... -
Synthetic promoter AAVs for cell-type specific expression in retinal cells
TypeBlog PostPublishedMarch 12, 2020, 1:15 p.m.... Roska’s lab at the Institute of Molecular and Clinical Ophthalmology Basel recently took a different ...Strategy #2 (ProB): Create promoters using phylogenetically conserved sequences that are upstream of the... -
Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene
TypeBlog PostPublishedApril 6, 2021, 12:30 p.m....given the data provided by the depositing lab, we call these “quality control (QC) issues.” We then ask..., Brigham MD, Sanjana NE, Zhang F (2017) Genome-scale CRISPR-Cas9 knockout and transcriptional activation... -
Hot Plasmids and Viral Preps - March 2021
TypeBlog PostPublishedMarch 23, 2021, 1:15 p.m....to visualize DNA, an editor targets DNA (via catalytically inactive dCas9) to modify chromatin. A fluorescent...INTRSECT project. Check out the new additions. Image calcium transients in PV interneurons with AAV-S5E2-GCaMP6f... -
Quickest Way to Deposit Plasmids: The Deposit Spreadsheet
TypeBlog PostPublishedMarch 8, 2017, 3:15 p.m....characters. A descriptive purpose focuses on the practical application of the plasmid and will help a scientist... Lysine (D123K), lowers biosensor affinity for calcium.” Primary Vector Type The vector type should... -
Cpf1 Update: Comparison to Cas9 and NgAgo
TypeBlog PostPublishedJuly 14, 2016, 2:30 p.m....Note: Cpf1 is also called Cas12a. In 2015, Feng Zhang’s lab characterized two Cpf1 nucleases, distant...should remove the PAM sequence. Many research and clinical applications require HDR over NHEJ, so comparing... -
High Complexity Golden Gate Assembly with NEB’s DAD
TypeBlog PostPublishedAug. 1, 2023, 1:15 p.m....search algorithm rather than pre-designed sets, so identical searches can return multiple answers. You might...might want to take advantage of their save and recall functions when you get results you like. NEBridge ... -
Design Tips for Prime Editing
TypeBlog PostPublishedJan. 23, 2025, 2:15 p.m.... gRNA (Anzalone et al., 2019), disrupting its canonical structure and Cas9 binding (Nishimasu et al., ...other bases in this location bind to Cas9 with canonical sgRNA structure (right). Created with BioRender.com... -
Progress Towards a PAM-Free CRISPR
TypeBlog PostPublishedMarch 7, 2024, 2:39 p.m.... for PAM-less editing Guides for CRISPR are typically selected by the ‘quality’ of the gRNA sequence...the Liu lab through phage-assisted evolution, specifically with the goal of generating Cas enzymes with... -
Plasmids 101: 5 factors to help you choose the right cloning method
TypeBlog PostPublishedAug. 21, 2018, 12:31 p.m....however, your cost per cloning reaction drops dramatically, as one tube of enzyme should be enough for ... guarantees, some cloning techniques do tip the scales in your favor. For starters, Golden Gate cloning... -
Transferable Skills: Negotiation
TypeBlog PostPublishedMarch 3, 2022, 2 p.m....contracts behind closed doors. Fig. 1: The mythical land of negotiations. You negotiate everyday...are negotiating for a specific time slot on the confocal microscope. You’re trying to get better images... -
Polymerase Chain Reaction Overview and Applications
TypeBlog PostPublishedAug. 10, 2021, 1:15 p.m....not as quantitative as other methods of PCR. Theoretically scientists should be able to determine the quantity...addition of a probe based fluorescent dye that intercalates with any dsDNA and the use of a fluorometer ...