... in your own lab. Table 1: Comparison Between Standard Restriction Cloning and Fast Cloning Requirement...ligated in vivo. Detailed FastCloning procedure To start the FastCloning process, the insert and vector are...efficiency cloning Now that you have a better understanding of the step by step procedure of FastCloning... 1. Celie, Patrick HN, Annabel HA Parret, and Anastassis Perrakis. "Recombinant cloning strategies for...

... in the mall. I took action, joined a gym, and started running on a treadmill. After being with the company...treatments, if available. As part of the team, I started adding mile after mile on my poor, poor legs. ...all seems lost in the War of Light, Look to the stars –                                     -for HOPE ...development. His areas of expertise include early stage drug discovery and pre-clinical development for ...

...of the manager is not to empower but to remove obstacles.” – Scott Adams, Dilbert cartoonist What makes...for example. So aside from a focus on removing obstacles, what makes a good manager? First, it takes an...consultation, advice and brainstorming. Employees must understand the part they play in the goals of the organization...their present positions By removing the primary obstacle of job dissatisfaction, the work can get done,...

...construct. DNA and gRNA are not to scale.   Let’s start with the simplest multiplexing situation: you only...number. First you generate four unique kanamycin-resistant plasmids, each containing a different gRNA target... all but one of your gRNAs into spectinomycin-resistant plasmids pX330S-2 to pX330S-(last gRNA number)...: Comparison of Multiplex Strategies including Standard PolIII-gRNA cassette, Csy4-cleavable cassette,...dicot plants, with a choice of either hygromycin or Basta selection. Yang Lab Single Transcript Multiplexing...

... are focusing on tagging for detection purposes (stay tuned for our affinity purification tag blog!). ...benefits – as discussed above – but there are a few instances when they should be used with caution or not at...Protein folding issues, steric hindrances, and destabilization can be brought about by the tag so it’s important...aggregation of your protein of interest!   Ready to start epitope tagging?   References and Resources References...

...field of science you are communicating from. For instance, a plumber, a physicist, an ambassador to Spain... an email without a recipient, so why would you start a communications project without an audience?  An... racial/ethnic background, patient or caregiver status, geographic location, sense of humor, media consumed...audience allows you to create common ground and understanding that you can use to get your message across....

...several years ago we partnered with Seqwell to establish a next-generation sequencing (NGS) QC platform...including only titration of AAV particles and a protein stain to assess purity. For labs that produce a small ...samples, serotypes, or both. Unfortunately, the standard AAV QC regimen does not address this.  Previous...analysis approach can be easily incorporated into standard QC regimens and will provide scientists with the...

...expression of XFPs. Any longer and the cells may start to show morphological abnormalities due to the excessive...Clearing large tissues - Fluorescent proteins are very stable in SeeDB2. However, SeeDB2 is not powerful enough...neuronal circuit tracing.     Marcus Leiwe is an assistant professor at Kyushu University. He has a particular...17972876. Kohl, Johannes, et al. "Ultrafast tissue staining with chemical tags." Proceedings of the National...

...detection assays, in vitro transduction, and silver stain approaches we show that there is no difference in...research community.    So, why are we so excited? For starters, we believe that this approach can be adapted ... of retroviral vectors: Enhancement in vector stability." Biotechnology and bioengineering 94.2 (2006)...PubMed PMID: 16528756. 2. Oh, Steve KW, et al. "Substantial overproduction of antibodies by applying osmotic...

...we are cloning an ORF, we want to clone from the start codon (ATG) to the stop codon (TGA, in this example...give specificity and to also be compatible with a standard PCR reaction (see PCR Video). Therefore, our Forward...reverse complement to get PCR amplification. We can start similarly, taking the final 18 bases of the ORF,...restriction cloning post. However, even more than with standard restriction cloning, it is essential that you ...

...insert. Since the standard ligation reaction is fairly quick, make sure you stay organized and prepare...TOPO cloning: 1. Create Your PCR Product: Design standard primers (no need to add unique restriction sites...Reaction into Competent Cells: You can use your standard lab protocol for this; however, you should reduce...

...Bandrowski et al., 2016). It’s also been well established that recombinant antibodies are excellent for... to contribute to the catalog. The Catalog Will Start With Neuroscience Antibodies and Grow From There...Email [email protected] to learn more or get started.  If you have suggestions about how to make the...Bradbury A, Plückthun A (2015) Reproducibility: Standardize antibodies used in research. Nature 518:27–29...

...usage has been limited by technical problems. For instance, it’s much more difficult to introduce a gene ...primary (hESCs) and cultured cells showed up to 70% resistance to diphtheria toxin, indicating a 70% frequency...Figure adapted from D’Astolfo et al. In certain instances, the potential benefits of nonviral transduction...protein purification (once a protocol has been established) may be less laborious than viral purification...

...a great start to a mentoring relationship, but if you don’t take this past the chatting stage you will...list available from this post. Use an activity to start a discussion. The Forced Choice Analysis is a fun...clarify and discuss values. The activity is just the start – it is the discussion after that will lead to learning...

...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...

...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...

...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...

...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 ...

... 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...

... (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...