We narrowed to 1,045 results for: sta
-
TypeBlog Post...Standard AAV vectors are generated with a producer cell line like the widely used 293T. During vector...glioblastoma). They also found that exo-AAVs were more resistant to neutralizing anti-AAV antibodies. This is particularly...thought to enhance the production of exosomes or stabilize their structure or their interaction with AAVs...areas that are difficult to reach, and are more resistant to inactivation through anti-AAV antibodies. ...
-
Unique and persistent IDs for improved reproducibility: Addgene now supports RRIDs and compact identifiers for all plasmids
TypeBlog Post...reagent is crucial for reproducibility because mistakes in this can lead to wasted time or retractions... the Addgene catalog. These IDs allow you to unmistakably identify a plasmid available through Addgene...all Addgene plasmids! In an effort to support standardized persistent identification of research reagents...Resource Identifiers (CURIEs) combined with well-established prefix resolution services such as Identifiers.org... -
Pushing the Limits of DNA Assembly
TypeBlog Post...is fine for its goal of creating a choice of standardized constructs and tools for the rational and simple...Golden Gate Assembly also allows the creation of standardized DNA constructs, but its use of Type IIS restriction...polymerase used in the amplification. This tool allows instant updates as fragments are added, deleted, or reordered... in biofuel generation, green chemistry and understanding the minimal genome. Many of these techniques... -
How Dry Ice Affects Viral Vector Sample pH & How to Avoid It
TypeBlog Post... (Fig. 1). However, once thawed, there was a substantial reduction (from 7.4 to 5.0) in the pH of samples...antibodies, because of effects of pH on protein stability (Murphy et al., 2013). However, the pH changes...infectivity of the virus. However, there was a substantial reduction in infectivity after 3 weeks of exposure...der Geer P, Manning MC, Fitchmun MI. Protein instability following transport or storage on dry ice. Nat... -
FPbase: A new community-editable fluorescent protein database
TypeBlog Post...range of characteristics (color, brightness, photostability, maturation, oligomerization), many of which...choosing a protein, we often direct them there as a starting point. However, there is a great deal more information...database relating to that specific protein (for instance: EGFP). Users can “favorite” certain proteins...of FPbase FPbase is still very much in the early stages of development, and there is plenty of room for... -
Revamp Your Lab Meetings With Creative Virtual Collaboration
TypeBlog Post...at odd times, new strategies can make socially distanced work more creative. An example could be making...signal the approach of a brain break, where everyone stands up, disappears from the video, and prepares a coffee...brainstorm on data in smaller groups, where for instance the presence of the PI in the main room inhibits...these weird times of shift working and social distancing. The take home message is the approaches/tools... -
Networking Follow-up: I Met a Person. Now What?
TypeBlog Post...give rise to a starfish shape. You can ask me about the different arms of this starfish sometime, but... plug-in called Maps. Figure 1: See a starfish? This is actually a map of Joanne Kamen's LinkedIn... it is completely impossible for me or anyone to stay in touch with all of their connections on a regular... -
What Good Citizenship Can Do for Reproducibility in Science
TypeBlog Post...different water filtration system, environmental stability in an incubator), but there’s a lot you can do...other researchers to help them troubleshoot and understand. All of this communication and sharing fits together...seriously. That’s why at Cell Press we developed STAR Methods to improve the structure and transparency... of checklists and guidelines to help authors understand what they are expected to do. All of that helps... -
Optimizing Donor DNA for Enhanced CRISPR Genome Editing
TypeBlog Post...efficiency of these editing reactions is low. Understanding the biological cause of this repair bias is ... reinforce a key scientific principle: that understanding a biological process in detail often suggests... efficiency of sequence replacement by HDR. Understanding Cas9 biophysics enhances HDR We first measured...released strand, 127bp in length, With 36bp on the PAM-distal side of the break and 91bp on the PAM-proximal ... -
How to Lead a Great Meeting
TypeBlog Post... the purpose of the meeting: Begin meetings by stating “The purpose of this meeting is to _________." ... what you need to accomplish to get to the next stage, and then adjourn. Define the tone of the meeting...template for project meetings. Team Meetings: For standing team meetings, provide agenda sections for each...meeting. Accountability: At your next meeting, start by checking in on the action items from last meeting... -
Fluorescent Proteins 101: Luciferase
TypeBlog Post...Imaging Improving Well-established High-throughput assays Improving Well-established Biological sources...cells. Luciferase assays have since become a gold standard in gene expression analysis and a luciferase gene...Phrixothrix hirtus), and a variety of marine crustaceans. Many of these luciferases have been subjected... -
Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools
TypeBlog Post...phenotypes than standard RNAi protocols without the need for mutant genetic backgrounds. The standard RNAi vector...biochemical and biophysical study involving protein stability or binding experiments. Potential experiments ...perform using the purified domains include protein stability, small molecule binding screens, peptide arrays... -
A Practical Approach to Choosing the B(right)est Fluorescent Protein
TypeBlog Post...at (and through) a couple of models. Before you start a new project with fluorescent proteins, the best...this series of posts are (i) brightness, (ii) photostability and (iii) aggregation tendency. Brightness...Without further information about properties like photostability or aggregation tendency, our choice would be...Goedhard and Marieke Mastop! Joachim Goedhart is an assistant professor at the Section of Molecular Cytology... -
Plasmids 101: Inducible Promoters
TypeBlog Post...promoters. Others are only active under specific circumstances. In this post, we’ll discuss inducible promoters...promoters, which can be switched from an OFF to an ON state, and how you might use these in your research. When...negative control. Positive inducible In the OFF state, the promoter is inactive because the activator ...transcription. Negative inducible In the OFF state, the promoter is inactive because a bound repressor... -
Antibodies 101: Buffers, Storage, and Conjugates
TypeBlog Post...reaction to understand if any of your buffer components could affect the reaction. For instance, sodium azide...either testing the antibody efficacy through a standard curve, another quick experiment, or using a different... to use, not sensitive to light in its inactive state, and cheap. Unless there’s a specific reason you... -
Viral Vectors 101: Gamma-Retroviral Packaging Systems
TypeBlog Post...This method utilizes a packaging cell line that stably expresses Gag-Pol and/or Env, eliminating the need...packaging cell line developed by Garry Nolan at Stanford (Swift, 2001) contains Gag-Pol and either an ecotropic...differentiated (non-dividing) cells. Both lentiviruses and standard γ-retroviruses use the Gag, Pol, and Env genes...provirus. 3’ LTR Terminates transcription started by 5' LTR by the addition of a poly A tract just... -
Viral Vectors 101: Viruses as Biological Tools
TypeBlog Post...infect cells and deliver cargo, it’s important to understand their differences to choose the best one for ...viruses Retroviruses are a family of viruses that stably integrate their genome into the host genome, setting... Type of Virus Pros Cons Retrovirus Stable Gene Expression Broad Tropism Only Infects ...virus safety and viral applications to help you start (safely!) thinking about the possibilities. Ready... -
Plasmids 101: Blue-white Screening
TypeBlog Post...for colonies that contain your plasmid is a great start when cloning, but how about being able to choose..., and remain the lovely whitish-cream color of standard E. coli. On the other hand, intact β-galactosidase...colonies that not only have your plasmid (antibiotic resistance), but also confirm those plasmids have your insert...plasmid-containing cells without using antibiotic resistance. These rely on cell lines susceptible to or dependent... -
Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins
TypeBlog Post...ratio, high sensitivity, responds quickly, is photostable, and is compatible with optogenetic control. ...proteins that bind to either the light- and dark- states of the photoswitchable cPYP, which changes in conformation... partners that bind to only the light- or dark- state of the photoswitchable proteins offers a powerful...binders (which bind to either the light- or dark- state of the photoswitchable proteins) fused to mVenus... -
Rosella: A Fluorescent pH-Biosensor for Studying Autophagy
TypeBlog Post...protein degradation due to other pathways, it is standard practice to compare degradation rates between ...identify because their contents can be at various stages of degradation. Additionally, electron microscopy...that affect autophagy (ATG) in yeast. Atg8p (“p” stands for protein) associates with autophagosomal membranes...green fluorescence. After 4 hours of nitrogen starvation to induce autophagy, red but not green fluorescence...